New heterocycle compounds and uses thereof for the prevention or treatment of diseases involving formation of amyloid plaques and/or where a dysfunction of the app metabolism occurs

ABSTRACT

The present invention relates to compounds having the following Formula (I) for use in the prevention and/or the treatment of diseases involving formation of amyloid plaques and/or where a dysfunction of the APP metabolism occurs.

The present invention concerns new heterocycle compounds as well as usesthereof, for the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs, such as Alzheimer's disease.

Alzheimer's disease is a progressive neurodegenerative disorder thatgradually damages the neurons in regions of the brain involved inmemory, learning and reasoning. It is characterized by extracellularaccumulation of amyloid β (Aβ) peptides forming amyloid plaques in thebrain. This accumulation of Aβ peptides in the brain has been shown toplay a key role in the development of Alzheimer's disease.

Alzheimer's disease affects brain functions, including short-term memoryloss, inability to reason, and the deterioration of language and theability to care for oneself. An estimated 3% of people between the agesof 65 and 74 have Alzheimer's disease, rising to about half those age 85and over.

WO 02/37118 discloses means for detecting pathological transformation ofthe amyloid protein precursor (APP) and their uses in diagnostic andtherapeutic applications in degenerative pathologies such as Alzheimer'sdisease.

WO 2006/051489 discloses the use of 1,4-bis(3-aminoalkyl)piperazinederivatives for the treatment of neurodegenerative diseases, whereinsaid derivatives could be used to rectify the metabolism of the amyloidprotein precursor (APP).

The aim of the invention is to provide new drugs for the treatment ofdiseases involving formation of amyloid plaques and/or where adysfunction of the APP metabolism occurs, such as Alzheimer's disease.

The aim of the invention is to provide new compounds having the propertyof rectifying the metabolism of APP, wherein said compounds have animproved effect on said metabolism over prior art compounds.

The present invention relates to compounds having the following formula(I):

wherein:

a represents a single bond or a double bond;

b represents a single bond or a double bond, provided that when a is asingle bond, then b is a double bond, and when a is a double bond, thenb is a single bond;

R₃ is absent when a is a double bond, or, when a is a single bond, R₃ ischosen from the groups consisting of: alkyl, cycloalkyl, aryl andheterocyclyl radicals, said alkyl, cycloalkyl, aryl, and heterocyclylradicals being possibly substituted;

R₄ is absent when b is a double bond, or, R₄ is H when b is a singlebond;

R₅ is chosen from the group consisting of:

-   -   H,    -   (C₁-C₁₂)alkyl,    -   OH, and    -   (C₁-C₁₂)alkoxy,

R₆ is chosen from the group consisting of:

-   -   H,    -   halogen, in particular F or Br,    -   CN,    -   OH,    -   (C₁-C₁₂)alkoxy,    -   (C₁-C₁₂)alkyl,    -   (C₆-C₃₀)aryl,    -   heteroaryl,    -   CO₂R, wherein R is an alkyl group comprising from 1 to 12 carbon        atoms,    -   NR_(a)R_(b), R_(a) and R_(b) being each independently chosen        from: H, alkyl, aralkyl, aryl, cycloalkyl, and heterocyclyl        groups, said alkyl, aralkyl, aryl, cycloalkyl, and heterocyclyl        groups being possibly substituted, or R_(a) and R_(b) forming        together with the nitrogen atom carrying them a possibly        substituted heterocyclyl group; and    -   a radical of formula: —X′—(CH₂)_(n)—NR′₁R′₂, wherein:        -   X′ is chosen from: CH₂, O, NH, CO, CH₂OCO, and NHCO;        -   n′ is 0, 1 or 2;        -   R′₁ and R′₂ are each independently chosen from H, alkyl,            aralkyl, aryl, cycloalkyl and heterocyclyl groups, said            alkyl, aralkyl, aryl, cycloalkyl, and heterocyclyl groups            being possibly substituted, or R′₁ and R′₂ may form together            with the nitrogen atom carrying them a possibly substituted            heterocyclyl group;

X is chosen from: CH₂, O, NH, CO, CH₂OCO, and NHCO;

n is 0, 1 or 2;

R₁ and R₂ are each independently chosen from H, alkyl, aralkyl, aryl,cycloalkyl and heterocyclyl groups, said alkyl, aralkyl, aryl,cycloalkyl and heterocyclyl groups being possibly substituted, or R₁ andR₂ may form together with the nitrogen atom carrying them a possiblysubstituted heterocyclyl group;

or its pharmaceutically acceptable salts, hydrates or hydrated salts orits polymorphic

crystalline structures, racemates, diastereisomers or enantiomers,

for use in the prevention and/or the treatment of diseases involvingformation

of amyloid plaques and/or where a dysfunction of the APP metabolismoccurs,

with the exclusion of the compounds having the following formula:

wherein:

—R₆ is H and R₈ is —CH₂—NEt₂;

—R₆ is OH and R₈ is chosen from: —CH₂—NEt₂, —CH₂—NHEt,

and

—R₆ is OMe and R₈ is —CH₂—NEt₂ or

The term “alkyl” means a saturated or unsaturated aliphatic hydrocarbongroup which may be straight or branched having 1 to 12 carbon atoms inthe chain. Preferred alkyl groups have 1 to 6 carbon atoms in the chain.“Branched” means that one or lower alkyl groups such as methyl, ethyl orpropyl are attached to a linear alkyl chain. <<Lower alkyl>> means 1 to4 carbon atoms in the chain which may be straight or branched. The alkylmay be substituted with one or more <<alkyl group substituants>> whichmay be the same or different, and include for instance halo, cycloalkyl,hydroxy, alkoxy, amino, acylamino, aroylamino, carboxy.

The term “halo” refers to the atoms of the group 17 of the periodictable (halogens) and includes in particular fluorine, chlorine, bromine,and iodine atom.

The terms “arylalkyl” or “aralkyl” refer to an alkyl moiety in which analkyl hydrogen atom is replaced by an aryl group. Examples of“arylalkyl” or “aralkyl” include benzyl and 9-fluorenyl groups.

The term “cycloalkyl” as employed herein includes saturated cyclic,bicyclic, tricyclic, or polycyclic hydrocarbon groups having 3 to 12carbons, wherein any ring atom capable of substitution may besubstituted by a substituent. Examples of cycloalkyl moieties include,but are not limited to, cyclohexyl and adamantyl.

The term “aryl” refers to an aromatic monocyclic, bicyclic, or tricyclichydrocarbon ring system, wherein any ring atom capable of substitutionmay be substituted by a substituent. Examples of aryl moieties include,but are not limited to, phenyl, naphthyl, and anthracenyl.

The term “heterocyclyl” refers to a nonaromatic 3-10 memberedmonocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ringsystem having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms ifbicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selectedfrom O, N, or S (e. g. , carbon atoms and 1-3, 1-6, or 1-9 heteroatomsof N, O, or S if monocyclic, bicyclic, or tricyclic, respectively),wherein any ring atom capable of substitution may be substituted by asubstituent.

The term “heteroaryl” refers to an aromatic 5-8 membered monocyclic,8-12 membered bicyclic, or 11-14 membered tricyclic ring system having1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms selected from O, N, or S (e.g. , carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S ifmonocyclic, bicyclic, or tricyclic, respectively), wherein any ring atomcapable of substitution may be substituted by a substituent.

The term “alkoxy” refers to an —O-alkyl radical.

The term “substituents” refers to a group “substituted” on an alkyl,cycloalkyl, heterocyclyl, aryl, aralkyl or heteroaryl group at any atomof that group. Suitable substituents include, without limitation, alkyl,alkenyl, alkynyl, alkoxy, halo, hydroxy, cyano, nitro, amino, SO₃H,sulfate, phosphate, perfluoroalkyl, perfluoroalkoxy, methylenedioxy,ethylenedioxy, carboxyl, oxo, thioxo, imino (alkyl, aryl, aralkyl),S(O)_(n) alkyl (where n is 0-2), S(O)_(n) aryl (where n is 0-2),S(O)_(n) heteroaryl (where n is 0-2), S(O)_(n) heterocyclyl (where n is0- 2), amine (mono-, di-, alkyl, cycloalkyl, aralkyl, heteroaralkyl, andcombinations thereof), ester (alkyl, aralkyl, heteroaralkyl), amide(mono-, di-, alkyl, aralkyl, heteroaralkyl, and combinations thereof),sulfonamide (mono-, di-, alkyl, aralkyl, heteroaralkyl, and combinationsthereof), unsubstituted aryl, unsubstituted heteroaryl, unsubstitutedheterocyclyl, and unsubstituted cycloalkyl.

The term “acyl” refers to an alkylcarbonyl, cycloalkylcarbonyl,arylcarbonyl, heterocyclylcarbonyl, or heteroarylcarbonyl substituent,any of which may be further substituted by substituents.

The term “oxo” refers to an oxygen atom, which forms a carbonyl whenattached to carbon, an N-oxide when attached to nitrogen, and asulfoxide or sulfone when attached to sulfur.

The term “alkenyl” as employed herein includes partially unsaturated,nonaromatic, hydrocarbon groups having 2 to 12 carbons, preferably 2 to6 carbons.

The term “alkylene” as employed herein refers to a divalent radicalcomprising from 1 to 12 carbon atoms, and preferably from 1 to 6 carbonatoms. Said radical may be represented by the formula (CH₂), wherein nis an integer varying from 1 to 12, and preferably from 1 to 6.

The compounds herein described may have asymmetric centers. Compounds ofthe present invention containing an asymmetrically substituted atom maybe isolated in optically active or racemic forms. It is well-known inthe art how to prepare optically active forms, such as by resolution ofracemic forms or by synthesis from optically active starting materials.All chiral, diastereomeric, racemic forms and all geometric isomericforms of a compound are intended, unless the stereochemistry or theisomeric form is specifically indicated.

“Pharmaceutically acceptable” means it is, within the scope of soundmedical judgment, suitable for use in contact with the cells of humansand lower animals without undue toxicity, irritation, allergic responseand the like, and are commensurate with a reasonable benefit/risk ratio.

The term “pharmaceutically acceptable salt” refers to salts which retainthe biological effectiveness and properties of the compounds of theinvention and which are not biologically or otherwise undesirable. Inmany cases, the compounds of the invention are capable of forming acidand/or base salts by virtue of the presence of amino and/or carboxylgroups or groups similar thereto. Pharmaceutically acceptable acidaddition salts may be prepared from inorganic and organic acids, whilepharmaceutically acceptable base addition salts can be prepared frominorganic and organic bases. For a review of pharmaceutically acceptablesalts see Berge, et al. ((1977) J. Pharm. Sd, vol. 66, 1). Theexpression “non-toxic pharmaceutically acceptable salts” refers tonon-toxic salts formed with nontoxic, pharmaceutically acceptableinorganic or organic acids or inorganic or organic bases. For example,the salts include those derived from inorganic acids such ashydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, andthe like, as well as salts prepared from organic acids such as acetic,propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric,ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic,benzoic, salicyclic, sulfanilic, fumaric, methanesulfonic, andtoluenesulfonic acid and the like.

In the context of the invention, the term “treating” or “treatment”, asused herein, means reversing, alleviating, inhibiting the progress of,or preventing the disorder or condition to which such term applies, orone or more symptoms of such disorder or condition.

While it is possible for the compounds of the invention having formula(I) to be administered alone it is preferred to present them aspharmaceutical compositions. The pharmaceutical compositions, both forveterinary and for human use, useful according to the present inventioncomprise at least one compound having formula (I) as above defined,together with one or more pharmaceutically acceptable carriers andoptionally other therapeutic ingredients.

In certain preferred embodiments, active ingredients necessary incombination therapy may be combined in a single pharmaceuticalcomposition for simultaneous administration.

As used herein, the term “pharmaceutically acceptable” and grammaticalvariations thereof, as they refer to compositions, carriers, diluentsand reagents, are used interchangeably and represent that the materialsare capable of administration to or upon a mammal without the productionof undesirable physiological effects such as nausea, dizziness, gastricupset and the like.

The preparation of a pharmacological composition that contains activeingredients dissolved or dispersed therein is well understood in the artand need not be limited based on formulation. Typically suchcompositions are prepared as injectables either as liquid solutions orsuspensions; however, solid forms suitable for solution, or suspensions,in liquid prior to use can also be prepared. The preparation can also beemulsified. In particular, the pharmaceutical compositions may beformulated in solid dosage form, for example capsules, tablets, pills,powders, dragees or granules.

The choice of vehicle and the content of active substance in the vehicleare generally determined in accordance with the solubility and chemicalproperties of the active compound, the particular mode of administrationand the provisions to be observed in pharmaceutical practice. Forexample, excipients such as lactose, sodium citrate, calcium carbonate,dicalcium phosphate and disintegrating agents such as starch, alginicacids and certain complex silicates combined with lubricants such asmagnesium stearate, sodium lauryl sulphate and talc may be used forpreparing tablets. To prepare a capsule, it is advantageous to uselactose and high molecular weight polyethylene glycols. When aqueoussuspensions are used they can contain emulsifying agents or agents whichfacilitate suspension. Diluents such as sucrose, ethanol, polyethyleneglycol, propylene glycol, glycerol and chloroform or mixtures thereofmay also be used.

The pharmaceutical compositions can be administered in a suitableformulation to humans and animals by topical or systemic administration,including oral, rectal, nasal, buccal, ocular, sublingual, transdermal,rectal, topical, vaginal, parenteral (including subcutaneous,intra-arterial, intramuscular, intravenous, intradermal, intrathecal andepidural), intracisternal and intraperitoneal. It will be appreciatedthat the preferred route may vary with for example the condition of therecipient.

The formulations can be prepared in unit dosage form by any of themethods well known in the art of pharmacy. Such methods include the stepof bringing into association the active ingredient with the carrierwhich constitutes one or more accessory ingredients. In general theformulations are prepared by uniformly and intimately bringing intoassociation the active ingredient with liquid carriers or finely dividedsolid carriers or both, and then, if necessary, shaping the product.

Total daily dose of the compounds of the invention administered to asubject in single or divided doses may be in amounts, for example, offrom about 0.001 to about 100 mg/kg body weight daily and preferably0.01 to 10 mg/kg/day. Dosage unit compositions may contain such amountsof such submultiples thereof as may be used to make up the daily dose.It will be understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including thebody weight, general health, sex, diet, time and route ofadministration, rates of absorption and excretion, combination withother drugs and the severity of the particular disease being treated.

Unexpectedly, the inventors have discovered that the compounds offormula (I) according to the present invention could be used to rectifythe metabolism of the Amyloid Protein Precursor (APP) on three essentialpoints:

1) increasing the carboxy-terminal fragments of APP (APP-CTFs) which allin common possess the last 50 aminoacids of APP, and especially thosehaving potential physiological activities, such as the α-stubs (APP-CTFsα) and the AICD (APP intra cellular domain) with neurotrophicproperties, 2) decreasing the production of the neurotoxic by-productsof APP, i.e. β-amyloid (Aβ) peptides, especially in their form 1-40 and1-42,

Therefore, the compounds of formula (I) as described herein are usefulin the treatment of all diseases where a dysfunction of the APPmetabolism occurs. These diseases also include pathologies involvingamyloid depot.

Among those diseases, one may in particular mention such as Alzheimer'sdisease, Lewy body disease, Down syndrome, amyloid angiopathy,Parkinson's disease, prion diseases, in particular Creutzfeldt-JakobDisease (CJD), amyotrophic lateral sclerosis (ALS), and frontotemporaldegeneration.

In formula (I) mentioned above, when R₆ is a radical—X′—(CH₂)_(n)—NR′₁R′₂, said radical may be identical to or differentfrom the radical —X—(CH₂)_(n)—NR₁R₂.

The present invention also relates to compounds of formula (I-1):

wherein R₁, R₂, R₅, R₆, X, and n are as defined above in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

The compounds of formula (I-1) are compounds of formula (I) wherein a isa double bond and b is a single bond, R₃ is none and R₄ is H.

The present invention also relates to the compounds of formula (I) forthe use as mentioned above, wherein R₅ is H.

The present invention also relates to the compounds of formula (I) forthe use as mentioned above, wherein R₁ and R₂ are ethyl.

The present invention also relates to the compounds of formula (I) forthe use as mentioned above, wherein R₁ and R₂ form together with N apyrrolidine group.

The present invention also relates to the compounds of formula (I) forthe use as mentioned above, wherein —NR₁R₂ is chosen from the groupconsisting of: —NEt₂, —NHtBu, pyrrolidinyl, morpholinyl, andN-methyl-piperazinyl.

The present invention also relates to the compounds of formula (I) forthe use as mentioned above, wherein R₆ is halogen, and preferably F.

The present invention also relates to compounds of formula (I-2):

wherein R₁, R₂, R₅, R₆, X, and n are as defined above in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

The present invention also relates to compounds of formula (I-3):

wherein R₁, R₂, R₅, and R₆ are as defined above in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

The compounds of formula (I-3) are compounds of formula (I-2) wherein Xis CH₂ and n is 0.

The present invention also relates to compounds of formula (I-4):

Preferably, the present invention relates to compounds of formula (I-4)as defined above, for use in the prevention and/or the treatment ofdiseases involving formation of amyloid plaques and/or where adysfunction of the APP metabolism occurs, wherein when R₆ is OH, then R₁and R₂ are not ethyl.

Preferably, the present invention relates to compounds of formula (I-4)as defined above, for use in the prevention and/or the treatment ofdiseases involving formation of amyloid plaques and/or where adysfunction of the APP metabolism occurs, wherein R₆ is halogen, andpreferably F, or alkyl.

A particular class of compounds which may be used in the presentinvention is constituted of compounds having the following formula(I-4-1):

wherein R₁ and R₂ are as defined above in formula (I),

and R₆ is chosen from: H, OH, and (C₁-C₁₂)alkoxy.

In formula (I-4-1), R₆ is preferably chosen from H, OH and OEt.

In formula (I-4-1), R₁ and R₂ are preferably ethyl or form together withN a pyrrolidine group.

Preferably, in formula (I-4-1), when R₆ is OH, then R₁ and R₂ are notethyl, or R₁ and R₂ do not form together with N a N-methyl-piperazinylgroup. Preferably, in formula (I-4-1), when R₆ is methoxy, then R₁ andR₂ are not ethyl.

Thus, a particularly preferred class of compounds which may be used inthe present invention is constituted of compounds having the followingformula (I-4-1-1):

wherein R₆ is as defined above, and is preferably chosen from H, OH andOEt.

Another particularly preferred class of compounds which may be used inthe present invention is constituted of compounds having the followingformula (I-4-1-2):

wherein R₆ is as defined above, and is preferably chosen from H, OH andOEt.

A particular class of compounds which may be used in the presentinvention is constituted of compounds having the following formula(I-4-2):

wherein R₁ and R₂ are as defined above in formula (I),

and R₆ is chosen from: (C₁-C₁₂)alkyl, (C₆-C₃₀)aryl, and heteroaryl, saidalkyl, aryl and heteroaryl being possibly substituted.

In formula (I-4-2), R₁ and R₂ are preferably ethyl or form together withN a pyrrolidine group.

In formula (I-4-2), R₆ is preferably Me, Et, furyl, thienyl, or apossibly substituted phenyl group. Among the substituents, thefollowings may be cited: alkyl such as CH₃ or tBu, perfluoroalkyl suchas CF₃, halogen such as F or Cl, alkoxy such as OCH₃, aryloxy such asOBn, alkylcarbonyl such as COCH₃, and perfluoroalkoxy such as OCF₃.

Thus, a particularly preferred class of compounds which may be used inthe present invention is constituted of compounds having the followingformula (I-4-2-1):

wherein R₆ is as defined above in formula (I-4-2).

Another particularly preferred class of compounds which may be used inthe present invention is constituted of compounds having the followingformula (I-4-2-2):

wherein R₆ is as defined above in formula (I-4-2).

Another particularly preferred class of compounds which may be used inthe present invention is constituted of compounds having the followingformula (I-4-2-3):

wherein R₁ and R₂ are as defined above in formula (I), and

R₇ is chosen from alkyl such as CH₃ or tBu, perfluoroalkyl such as CF₃,halogen such as F or Cl, alkoxy such as OCH₃, aryloxy such as OBn,alkylcarbonyl such as COCH₃, and perfluoroalkoxy such as OCF₃.

A particular class of compounds which may be used in the presentinvention is constituted of compounds having the following formula(I-4-3):

wherein R_(a), R_(b), R₁ and R₂ are as defined above in formula (I).

In formula (I-4-3), R₁ and R₂ are preferably ethyl or form together withN a pyrrolidine group.

Preferably, in formula (I-4-3), R_(a) and R_(b) form together with thenitrogen atom carrying them a morpholinyl or a N-methyl-piperazinylgroup. A particular class of compounds which may be used in the presentinvention is constituted of compounds having the following formula(I-4-4):

wherein R₁ and R₂ are as defined above in formula (I).

Preferably, in formula (I-4-4), R₆ is Br or F, and most preferably F.

Preferably, in formula (I-4-4), R₁ and R₂ are preferably ethyl or formtogether with N a pyrrolidine group.

The present invention also relates to compounds of formula (I-3-1):

wherein R₁, R₂ and R₆ are as defined above in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

Preferably, in formula (I-3-1), R₆ is OH.

The present invention also relates to compounds of formula (I-3-2):

wherein R₁ and R₂ are as defined above in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

Preferably, in formula (I-3-1) or (I-3-2), —NR₁R₂ is chosen from thegroup consisting of: —NEt₂, —NHtBu, pyrrolidinyl, morpholinyl, andN-methyl-piperazinyl.

The present invention also relates to compounds of formula (I-5):

wherein R₁, R₂, R₅, R_(a) and R_(b) are as defined in formula (I),

R₅ being preferably H or CH₃,

R_(a) and R_(b) preferably forming with the nitrogen atom carrying thema possibly substituted heterocyclyl group, and more preferably R_(a) andR_(b) forming with the nitrogen atom carrying them a N-methylpiperazineor a morpholine group, for use in the prevention and/or the treatment ofdiseases involving formation of amyloid plaques and/or where adysfunction of the APP metabolism occurs.

According to a preferred embodiment, in above formula (I-5), R₅ is H.

According to another preferred embodiment, in above formula (I-5), R₅ isCH₃.

The present invention also relates to compounds of formula (I-6):

wherein R₁, R₂ and R₅ are as defined in formula (I),

and wherein X is CO or CH₂)

R₅ being preferably H or OH, and more preferably H, for use in theprevention and/or the treatment of diseases involving formation ofamyloid plaques and/or where a dysfunction of the APP metabolism occurs.

According to a preferred embodiment, in above formula (I-6), R₅ is H.

According to another preferred embodiment, in above formula (I-6), R₅ isOH.

A particular class of compounds which may be used in the presentinvention is constituted of compounds having the following formula(I-6-1):

Compounds of formula (I-6-1) correspond to compounds having formula(I-6) wherein R₅ is H and X is CO.

A particular class of compounds which may be used in the presentinvention is constituted of compounds having the following formula(I-6-2):

Compounds of formula (I-6-2) correspond to compounds having formula(I-6) wherein R₅ is OH and X is CH₂.

Preferably, in formula (I-6-2), R₁ and R₂ form together with thenitrogen atom carrying them a pyrrolidinyl, a N-methylpiperazinyl or amorpholinyl group.

A particular class of compounds which may be used in the presentinvention is constituted of compounds having the following formula(I-7):

wherein R₁ and R₂ are as defined in formula (I).

The present invention also relates to compounds of formula (I-8):

wherein R₁ and R₂ are as defined in formula (I), and form together withthe nitrogen atom carrying them a heterocycle,

and R₆ is as defined in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

The present invention also relates to compounds of formula (I-9):

wherein R₁ and R₂ are as defined in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

The present invention also relates to compounds of formula (1-10):

wherein R₁ and R₂ are as defined in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

The present invention also relates to compounds of formula (I-11):

wherein R₁ and R₂ are as defined in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

The present invention also relates to compounds of formula (I-12):

wherein R₁ and R₂ are as defined in formula (I),

for use in the prevention and/or the treatment of diseases involvingformation of amyloid plaques and/or where a dysfunction of the APPmetabolism occurs.

Some of the compounds of formula (I) are already known. Indeed, variouscompounds of formula (I) are useful for their antimalarial properties.

In particular, compounds of formula (I-4) are disclosed in (a)<<Synthesis and Antimalarial Activity of New Analogues of Amodiaquine>>,Delarue-Cochin, S.; Paunescu, E.; Maes, L.; Mouray, E.; Sergheraert, C.;Grellier, P.; Melnyk, P. European Journal of Medicinal Chemistry 2008,43, 252-260, (b) <<Replacement of 4′-hydroxy group of Amodiaquine andAmopyroquine by aromatic and aliphatic substituants: Synthesis andAntimalarial Activity>>, Paunescu, E.; Susplugas, S.;

Boll, E.; Varga, R.; Mouray, E.; Grosu, I.; Grellier, P.; Melnyk, P.,ChemMedChem. 2009, 4 (4), 549-561; (c) <<Suzuki coupling reaction as thekey step for the synthesis of 4′-substituted analogues of Amodiaquine>>,Paunescu, E.; Matuszak, N.; Melnyk, P., Tetrahedron 2007, 63,12791-12810; and (d) <<Synthesis and Antimalarial Activity of New AminoAnalogues of Amodiaquine and Amopyroquine>>, Paunescu, E.; Susplugas,S.; Boll, E.; Vargas, R. A.; Mouray, E.; Grellier, P.; Melnyk, P.,Medicinal Chemistry 2008, 4 (5), 407-425.

In particular, compounds of formula (I-9) are disclosed in <<Synthesisand in vitro and in vivo Antimalarial Activity of New4-aminoquinolines>>, Delarue-Cochin, S.; Girault, S.; Maes, L.;Debreu-Fontaine, M.-A.; Labaeïd, M.; Grellier, P.; Sergheraert, C.,Journal of Medicinal Chemistry 2001, 44, 2827-2833.

In particular, compounds of formula (I-10) are disclosed in <<Synthesisand in vitro and in vivo Antimalarial Activity of New4-aminoquinolines>>, Delarue-Cochin, S.; Girault, S.; Maes, L.;Debreu-Fontaine, M.-A.; Labaeïd, M.; Grellier, P.; Sergheraert, C.,Journal of Medicinal Chemistry 2001, 44, 2827-2833; and in <<Synthesisand Antimalarial Activity of New Analogues of Amodiaquine>>,Delarue-Cochin, S.; Paunescu, E.; Maes, L.; Mouray, E.; Sergheraert, C.;Grellier, P.; Melnyk, P. European Journal of Medicinal Chemistry 2008,43, 252-260.

In particular, compounds of formulae (I-11) and (I-12) are disclosed in<<Synthesis and Antimalarial Activity of Carbamate and Amide Derivativesof 4-anilinoquinoline>>, Delarue-Cochin, S.; Grellier, P.; Maes, L.;Mouray, E.; Sergheraert, C.; Melnyk, P., European Journal of MedicinalChemistry. 2008, 43, 2045-2055.

Some of the compounds of formula (1) are new compounds: compounds havingformulae (I-5), (I-6) wherein R₅=H, (I-7), and (I-8) as defined abovehave never been disclosed.

The present invention also relates to compounds having following formula(I-5-1):

wherein R₁, R₂, R_(a) and R_(b) are as defined in formula (I),

R_(a) and R_(b) preferably forming with the nitrogen atom carrying thema possibly substituted heterocyclyl group, and more preferably R_(a) andR_(b) forming with the nitrogen atom carrying them a N-methylpiperazineor a morpholine group.

Preferably, in formula (I-5-1), R₁ and R₂ form together with thenitrogen atom carrying them a N-methylpiperazinyl group or R₁ and R₂ areethyl.

The present invention also relates to compounds having following formula(I-5-2) or (I-5-3):

wherein R₁ and R₂ are as defined in formula (I).

The present invention also relates to compounds having following formula(I-5-4):

wherein R₁, R₂, R_(a) and R_(b) are as defined in formula (I),

R_(a) and R_(b) preferably forming with the nitrogen atom carrying thema possibly substituted heterocyclyl group, and more preferably R_(a) andR_(b) forming with the nitrogen atom carrying them a N-methylpiperazineor a morpholine group.

Preferred classes of compounds consist of compounds having formulae(I-6-1) or (I-6-2) as follows:

wherein R₁ and R₂ are as defined in formula (I).

Preferred classes of compounds consist of compounds having formulae(I-8-1) or (I-8-2) as follows:

R₆ being as defined in formula (I).

Another preferred class of compounds consists of compounds havingformula (I-8-3) as follows

R₁ and R₂ being as defined in formula (I).

The present invention also relates to compounds of formula (I-13):

a, b, R₃, R₁ and R₄ being as defined in formula (I).

A preferred class of compounds of the invention consists of compoundshaving formula (I-13-1):

R₁ being as defined in formula (I),

R₁ representing preferably H or

The present invention also relates to a pharmaceutical compositioncomprising the compounds having formulae (I-5), (I-5-1), (I-5-2),(I-5-3), (I-5-4), (I-6), (I-6-1), (I-6-2), (I-8), (I-8-1), (I-8-2),(I-8-3), (I-13), and (I-13-1) as defined above, in association with apharmaceutically acceptable vehicle.

The compounds of formula (I) have the property of decreasing thesecretion of the Aβ peptide (80% at 1 μM, and almost no secretion at 5μM), and of increasing the production of APP-CTF alpha stubs and AICD(×30-40 at 5 μM).

The present invention also relates to a method for the preparation ofcompounds having formula (I-8-3) as defined above, said methodcomprising the following steps:

a) a step of reacting 3,5-dibromobenzene with a compound having formulaHNR₁R₂, R₁ and R₂ being as defined in formula (I), to obtain a compoundhaving the following formula (I-8-3-1):

b) a step of hydrogenating the compound of formula (I-8-3-1) to obtain acompound having the following formula (I-8-3-2):

c) and a step of reacting the compound of formula (I-8-3-2) with4,7-dichloroquinoline to obtain the compound of formula (I-8-3), saidstep being possibly followed by a step of recovering said compound.

Preferably, 3,5-dibromobenzene is prepared from2,6-dibromo-4-nitroanaline, which is in particular prepared fromp-nitroaniline (Sheperd et al., 1947).

Preferably, step a) is carried out in the presence of BINAP(2,2′-bis(diphenylphosphino)-1,1′-binaphthyl), Cs₂CO₃, Pd₂dba₃(tris(dibenzylidene-acetone)dipalladium(0)) and dry 1,4-dioxane.

Preferably, hydrogenation step b) is carried out in the presence ofammonium formate and Pd/C in EtOH.

Preferably, step c) is carried out in the presence of BINAP, Cs₂CO₃,Pd₂dba₃ and dry 1,4-dioxane.

EXPERIMENTAL PART A—Synthesis of the New Compounds General Procedure A:AlMe₃ Promoted Amide Formation

A solution of AlMe₃ (2M in toluene, n eq) was added dropwise to asolution of the appropriate amine (n eq) in anhydrous DCM at 0° C. Thereaction mixture was stirred for 30 min and then allowed to warm up toroom temperature. 1,3-Dimethyl5-[(7-chloroquinolin-4-yl)amino]benzene-1,3-dicarboxylate 1 was addedportionwise and the reaction mixture was refluxed for 24 h. The mixturewas then poured in crushed ice and evaporated. The residue was suspendedin a DCM/methanol mixture and the precipitate was removed by filtration.The filtrate was evaporated and purified by TLC.

Example 1 Preparation of7-chloro-N-[4-(4-methylpiperazin-1-yl)-3-[(4-methylpiperazin-1-yl)methyl]phenyl]quinolin-4-amine(compound 55) Step A: [2-(4-Methylpiperazin-1-yl)-5-nitrophenyl]methanol

2-Fluoro-5-nitrobenzyl alcohol (1 g, 5.84 mmol) and N-methylpiperazine(1.29 mL, 2 eq) were heated at 130° C. for 1 h. The reaction mixture wasthen diluted with 25 mL of THF and washed with a saturated aqueoussolution of Na₂CO₃ and then with brine. The organic layer was dried overNa₂SO₄, filtered and evaporated. The residue was thoroughly washed withpentane and filtered off to yield expected compound as an orange powder(1.38 g, 94% yield). m/z (ESI) 252.1 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.46; (d, J=2.8 Hz, 1H), 8.18; (dd, J=8.9 and2.9 Hz, 1H), 7.26; (d, J=8.90 Hz, 1H), 4.75; (s, 2H), 3.20-3.10; (m,4H), 2.75-2.65; (m, 4H), 2.44 (s, 3H); ¹³C NMR (75 MHz, MeOH-d₄) δ156.5;(C), 143.4; (C), 136.6; (C), 124.1; (CH), 123.6; (CH), 119.0; (CH),59.5; (CH₂), 55.2; (CH₂), 51.7; (CH₂), 45.2; (CH₃).

Step B:1-methyl-4-{2-[(4-methylpiperazin-1-yl)methyl]-4-nitrophenyl}piperazine

To [2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methanol (0.150 g, 0.60mmol) in 5 mL of THF at 0° C. was added thionyl chloride (0.22 mL, 5eq). The reaction mixture was refluxed for 45 minutes and then thesolvent and the excess of thionyl chloride were evaporated. The residuewas suspended in 5 mL of acetonitrile and 1-methylpiperazine (0.33 mL, 5eq) was added at 0° C. The solution was stirred overnight at roomtemperature, evaporated and the residue was dissolved in ethyl acetateand washed with a saturated aqueous solution of Na₂CO₃. The aqueouslayer was extracted with ethyl acetate and then the combined organiclayers were dried over Na₂SO₄, filtered and evaporated. The residue wasthoroughly washed with pentane and filtered off to yield expectedcompound as an orange powder (0.185 g, 92% yield).

¹H NMR (300 MHz, CDCl₃) δ8.23; (d, J=2.8 Hz, 1H), 7.96; (dd, J=8.9 and2.8 Hz, 1H), 6.94; (d, J=8.9 Hz, 1H), 3.40; (s, 2H), 3.10-3.00; (m, 4H),2.60-2.25; (m, 12H), 2.27; (s, 3H), 2.19; (s, 3H).

Step C:4-(4-methylpiperazin-1-yl)-3-[(4-methylpiperazin-1-yl)methyl]aniline

A mixture of SnCl₂ (0.398 g, 2.1 mmol), 1.6 mL of HCl 1M and 5 mL of THFwas added to a solution of1-methyl-4-{2-[(4-methylpiperazin-1-yl)methyl]-4-nitrophenyl}piperazine(0.175 g, 2.5 eq) in 15 mL of THF. The reaction mixture was refluxed for3 h and allowed to cool to room temperature. A saturated aqueoussolution of Na₂CO₃ was added. The aqueous layer was extracted with DCM.The combined organic layers were dried over Na₂SO₄, filtered andevaporated. The residue was purified by flash chromatography(DCM/MeOH/NH₄OH//8/2/0.1) to yield expected compound as a brown oil (100mg, 63% yield).

¹H NMR (300 MHz, CDCl₃) δ6.89; (d, J=8.4 Hz, 1H), 6.71; (d, J=2.7 Hz,1H), 6.51; (dd, J=8.9 and 3.0 Hz, 1H), 3.45; (s, 2H), 2.83; (t, J=4.8Hz, 4H), 2.60-2.30; (m, 12H), 2.28; (s, 3H), 2.22; (s, 3H); ¹³C NMR (75MHz, CDCl₃) δ144.2; (C), 142.5; (C), 134.7; (C), 121.4; (CH), 117.4;(CH), 114.4; (CH), 57.3; (CH₂), 55.9; (CH₂), 55.3; (CH₂), 53.1; (CH₂),53.1; (CH₂), 46.2; (CH₃), 46.1; (CH₃).

Step D:7-chloro-N-[4-(4-methylpiperazin-1-yl)-3-[(4-methylpiperazin-1-yl)methyl]phenyl]quinolin-4-amine

4-(4-methylpiperazin-1-yl)-3-[(4-methylpiperazin-1-yl)methyl]aniline (68mg, 0.22 mmol) and 4,7-dichloroquinoline (44 mg, 1 eq) were refluxedovernight in 10 mL of acetonitrile with 0.66 mL of HCl 1M. The reactionmixture was then evaporated and purified by preparative thin-layerchromatography (DCM/MeOH/NH₄O/18/210.1) to yield expected compound as apale yellow solid (49 mg, 48% yield). m/z (ESI) 465.2 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.37; (d, J=5.6 Hz, 1H), 8.31; (d, J=9.1 Hz,1H), 7.88; (d, J=2.1 Hz, 1H), 7.53; (dd, J=9.1 and 2.1 Hz, 1H), 7.47;(s, 1H), 7.33-7.26; (m, 2H), 6.84; (d, J=5.6 Hz, 1H), 3.68; (s, 2H),3.76; (s, 2H), 3.12-3.00; (m, 4H), 2.85-2.45; (m, 12H), 2.42; (s, 3H),2.33; (s, 3H); ¹³C NMR (75 MHz, MeOH-d₄) δ154.1; (C), 149.8; (C), 145.8;(CH), 142.5; (C), 138.8; (C), 134.5; (C), 134.5; (C), 127.6; (CH),127.3; (CH), 125.1; (CH), 125.1; (CH), 122.2; (CH), 121.7; (CH), 116.9;(C), 100.8; (CH), 56.5; (CH₂), 54.6; (CH₂), 53.9; (CH₂), 50.4; (CH₂),50.0; (CH₂), 42.8; (CH₃), 42.6; (CH₃).

Example 2 Preparation of N-{3-[(tert-butylamino)methyl]-4-(4-methylpiperazin-1-yl)phenyl}-7-chloroquinolin-4-amine (compound 56) Step A:tert-butyl({[2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methyl})amine

To [2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methanol prepared in step Aof example 1 (0.150 g, 0.60 mmol) in 5 mL of THF at 0° C. was addedthionyl chloride (0.22 mL, 5 eq). The reaction mixture was refluxed for45 minutes and then the solvent and the excess of thionyl chloride wereevaporated. The residue was suspended in 5 mL of acetonitrile andtert-butylamine (0.62 mL, 10 eq) was added at 0° C. The solution wasstirred 48 h at room temperature, evaporated and the residue wasdissolved in ethyl acetate and washed with a saturated aqueous solutionof Na₂CO₃. The aqueous layer was extracted with ethyl acetate and thenthe combined organic layers were dried over Na₂SO₄, filtered andevaporated. The residue was thoroughly washed with pentane and filteredoff to yield expected compound as an orange powder (0.192 g,quantitative yield) m/z (ESI) 307.2 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ8.19; (d, J=2.8 Hz, 1H), 7.99; (dd, J=8.9 and2.8 Hz, 1H), 6.96; (d, J=8.9 Hz, 1H), 3.68; (s, 2H), 3.09; (t, J=4.6 Hz,4H), 2.60-2.45; (m, 4H), 2.31; (s, 3H), 1.11; (s, 9H); ¹³C NMR (75 MHz,CDCl₃) δ157.1; (C), 142.7; (C), 135.7; (C), 126.0 ;(CH), 123.3; (CH),118.7; (CH), 55.3; (CH₂), 51.8; (CH₂), 50.7; (C), 46.1; (CH₃), 42.7;(CH₂), 29.1; (CH₃).

Step B: 3-[(tert-butylamino)methyl]-4-(4-methylpiperazin-1-yl)aniline

A mixture of SnCl₂ (0.364 g, 4 eq), 1.44 mL of HCl 1M and 5 mL of THFwas added to a solution oftert-butyl({[2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methyl})amine(0.147 g, 0.48 mmol) in 20 mL of THF. The reaction mixture was refluxedfor 3 h and allowed to cool to room temperature. A saturated aqueoussolution of Na₂CO₃ was added. The aqueous layer was extracted with DCM.The combined organic layers were dried over Na₂SO₄, filtered andevaporated. The residue was purified by flash chromatography(DCM/MeOH/NH₄OH//85/15/1) to yield expected compound as a brown oil (65mg, 49% yield).

¹H NMR (300 MHz, CDCl₃) δ6.87; (d, J=8.4 Hz, 1H), 6.55; (d, J=2.7 Hz,1H), 6.43; (dd, J=8.4 and 2.7 Hz, 1H), 3.57; (s, 2H), 2.79; (t, J=4.5Hz, 4H), 2.44; (br, 4H), 2.24; (s, 3H), 1.09; (s, 9H); ¹³C NMR (75 MHz,CDCl₃) δ143.3; (C), 142.8; (C), 136.8; (C), 121.8; (CH), 117.0; (CH),114.3; (CH), 55.8; (CH₂), 53.0; (CH₂), 50.7; (C), 46.0; (CH₃), 43.6;(CH₂), 28.8; (CH₃).

Step C:N-{3-[(tert-butylamino)methyl]-4-(4-methylpiperazin-1-yl)phenyl}-7-chloroquinolin-4-amine

3-[(tert-butylamino)methyl]-4-(4-methylpiperazin-1-yl)aniline (60 mg,0.22 mmol) and 4,7-dichloroquinoline (43 mg, 1 eq) were refluxedovernight in 10 mL of acetonitrile with 0.43 mL of HCl 1M. The reactionmixture was then evaporated and purified by preparative thin-layerchromatography (DCM/MeOH/NH₄OH/18/210.1) to yield expected compound as apale yellow solid (45 mg, 47% yield). m/z (ESI) 438.3 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.36; (d, J=5.6 Hz, 1H), 8.27; (d, J=9.1 Hz,1H), 7.85; (d, J=2.1 Hz, 1H), 7.49; (dd, J=9.1 and 2.1 Hz, 1H),7.39-7.24; (m, 3H), 6.90; (d, J=5.6 Hz, 1H), 3.82; (s, 2H), 3.02; (t,J=4.7 Hz, 4H), 2.67; (br, 4H), 2.39; (s, 3H), 1.24; (s, 9H); ¹³C NMR (75MHz, MeOH-d₄) δ151.5; (CH), 150.4; (C), 149.2; (C), 148.9; (C), 137.2;(C), 136.7; (C), 135.7; (C), 126.8; (CH), 126.1; (CH), 125.6; (CH),123.8; (CH), 123.7; (CH), 122.2; (CH), 118.4; (C), 101.5; (CH), 55.8;(CH₂), 52.7; (CH₂), 51.1; (C), 45.3; (CH₃), 43.4; (CH₂), 27.9; (CH₃).

Example 3 Preparation of7-chloro-N-[4-(4-methylpiperazin-1-yl)-3-(morpholin-4-ylmethyl)phenyl]quinolin-4-amine(compound 57) Step A: [5-amino-2-(4-methylpiperazin-1-yl)phenyl]methanol

[2-(4-Methylpiperazin-1-yl)-5-nitrophenyl]methanol prepared in step A ofexample 1 (1.00 g, 3.98 mmol) was hydrogenated using ammonium formate(2.51 g, 10 eq) and Pd/C (10% Pd, 0.43 mg, 0.1 eq) in 50 mL of EtOH. Themixture was stirred overnight at room temperature and then filteredthrough a celite pad. The filtrate was evaporated and the residue wasdissolved in DCM and washed with an saturated aqueous solution ofNa₂CO₃. The aqueous layer was extracted with DCM. The combined organiclayers were dried over Na₂SO₄, filtered and evaporated to yield expectedcompound as a pale yellow oil (0.85 g, 97% yield).

¹H NMR (300 MHz, MeOH-d₄) δ7.00; (d, J=8.4 Hz, 1H), 6.78; (d, J=2.7 Hz,1H), 6.64; (dd, J=8.7 and 2.7 Hz, 1H), 4.65; (s, 2H), 2.95-2.85; (m,4H), 2.70-2.50 (m, 4H), 2.34; (s, 3H); ¹³C NMR (75 MHz, MeOH-d₄) δ144.5;(C), 142.2; (C), 137.1; (C), 121.2; (CH), 115.7; (CH), 115.2; (CH),61.2; (CH₂), 55.8; (CH₂), 52.7; (CH₂), 45.2; (CH₃).

Step B:{5-[(7-chloroquinolin-4-yl)amino]-2-(4-methylpiperazin-1-yl)phenyl}methanol

[5-amino-2-(4-methylpiperazin-1-yl)phenyl]methanol (100 mg, 0.45 mmol)and 4,7-dichloroquinoline (94 mg, 1.1 eq) were refluxed in 1 mL ofn-pentanol overnight. The reaction mixture was then evaporated andpurified by flash chromatography (DCM/MeOH/NH₄OH//8/2/0.1) to yieldexpected compound as a pale brown solid (143 mg, 83% yield). m/z (ESI)383.1 [M+H]⁺;

¹H NMR (300 MHz, MeOH-d₄) δ8.42-8.34; (m, 2H), 7.89; (d, J=2.0 Hz, 1H),7.58; (dd, J=9.1 and 2.1 Hz, 1H), 7.55-7.53; (m, 1H), 7.35-7.28; (m,2H), 6.89; (d, J=6.1 Hz, 1 H), 4.78; (s, 2H), 3.20-3.02; (m, 8H), 2.69;(s, 3H); ¹³ NMR (75 MHz, MeOH-d₄) δ152.1; (C), 149.3; (CH), 148.0; (C),146.7; (C), 138.3; (C), 137.0; (C), 135.7; (C), 126.4; (CH), 124.9;(CH), 124.8; (CH), 124.2; (CH), 124.1; (CH), 121.1; (CH), 117.9; (C),101.2; (CH), 59.9; (CH₂), 55.0; (CH₂), 51.4; (CH₂), 44.0; (CH₃).

Step C:7-chloro-N-[4-(4-methylpiperazin-1-yl)-3-(morpholin-4-ylmethyl)phenyl]quinolin-4-amine

To{5-[(7-chloroquinolin-4-yl)amino]-2-(4-methylpiperazin-1-yl)phenyl}methanol(0.300 g, 0.78 mmol) in 6 mL of DMF at 0° C. was added thionyl chloride(0.28 mL, 5 eq). The reaction mixture was warmed up to room temperatureand stirred for 2 h. The solvent was evaporated. The residue wassuspended in 6 mL of DMF and morpholine (1.39 mL, 20 eq) was added at 0°C. The solution was stirred overnight at room temperature, evaporatedand the residue was purified by flash chromatography(DCM/MeOH/NH₄OH//9/1/0.1) to yield expected compound as a pale yellowsolid (238 mg, 67% yield). m/z (ESI) 452.2 [M+H]⁺;

¹H NMR (300 MHz, MeOH-d₄) δ8.51 (d, J=5.3 Hz, 1H), 8.00; (d, J=2.0 Hz,1H), 7.89; (d, J=9.0 Hz, 1H), 7.47-7.38; (m, 2H), 7.23-7.13; (m, 2H),6.88-6.82; (m, 2H), 3.72-3.65; (m, 4H), 3.57; (s, 4H), 3.07-2.98; (m,4H), 2.70-2.47; (m, 8H), 2.36; (s, 3H)

Example 4 Preparation of7-chloro-N-[3-({[2-(dimethylamino)ethyl](methyl)amino}methyl)-4-(4-methylpiperazin-1-yl)phenyl]quinolin-4-amine(compound 58)

To{5-[(7-chloroquinolin-4-yl)amino]-2-(4-methylpiperazin-1-yl)phenyl}methanolprepared in step B of example 3 (0.100 g, 0.26 mmol) in 2 mL of DMF at0° C. was added thionyl chloride (0.19 mL, 10 eq). The reaction mixturewas warmed up to room temperature and stirred for 2 h. The solvent wasevaporated. The residue was suspended in 2 mL of THF andN,N,N′-trimethylethylenediamine (0.33 mL, 10 eq) was added at 0° C. Thesolution was stirred overnight at room temperature, evaporated and theresidue was purified by preparative thin-layer chromatography(DCM/MeOH/NH₄OH//8/2/0.1). 232 mg of a yellow oil was obtained. Theresidue was dissolved in a minimum amount of methanol and precipitatedwith acetone to yield expected compound as a pale yellow solid (80 mg,66% yield). m/z (MALDI-TOF) 467.3 [M+H]⁺;

¹H NMR (300 MHz, D₂O) δ8.58; (d, J=9.1 Hz, 1H), 8.53; (d, J=7.1 Hz, 1H),8.16; (d, J=2.0 Hz, 1H), 7.88-8.00; (m, 4H), 7.11; (d, J=7.1 Hz, 1H),4.98; (s, 2H), 4.10-4.20; (m, 2H), 3.83-4.00; (m, 4H), 3.63-3.80; (m,2H), 3.45-3.60; (m, 4H), 3.42; (s, 6H), 3.24; (s, 3H), 3.07; (s, 3H);¹³C NMR (75 MHz, D₂O) δ158.0; (C), 153.9; (C), 145.1; (CH), 142.3; (C),141.1; (C), 137.2; (C), 134.0; (CH), 132.3; (CH), 130.5; (CH), 127.3;(CH), 126.9; (C), 126.7; (CH), 121.8; (CH), 118.3; (C), 102.8; (CH),66.3; (CH₂), 61.7; (CH₂), 56.1; (CH₂), 52.2; (CH₃), 45.7; (CH₃), 44.2;(CH₂), 35.9; (CH₃).

Example 5 Preparation of7-Chloro-N-[3-methyl-4-(4-methylpiperazin-1-yl)-5-(pyrrolidin-1-ylmethyl)phenyl]quinolin-4-amine(compound 59) Step A :2-[(2-Bromo-3-methyl-5-nitrophenyl)methyl]-2,3-dihydro-1H-isoindole-1,3-dione

N-(Hydroxymethyl)phtalimide (2.36 g, 13.3 mmol) was dissolved in 20 mLof triflic acid at 0° C. The mixture was stirred for 20 minutes and then2-bromo-5-nitrotoluene (2.88 g, 1 eq) was added. This solution wasallowed to warm to room temperature and was stirred for 18 h. Themixture was poured slowly into 300 mL of ice-cold water. The aqueouslayer was extracted with DCM. The combined organic layers were washedwith 100 mL of water, dried over Na₂SO₄, filtered and evaporated toyield expected compound as a white solid (4.71 g, 94% yield).

¹H NMR (300 MHz, CDCl₃) δ7.97; (d, 1H, J=2.6 Hz), 7.84; (m, 2H), 7.71;(m, 3H), 4.95; (s, 2H), 2.48; (s, 3H); ¹³C NMR (75 MHz, CDCl₃) δ167.7;(C), 146.7; (C), 140.9; (C), 137.3; (C), 134.5; (CH), 132.3; (C), 131.7;(C), 124.1; (CH), 123.8; (CH), 119.9; (CH), 42.3; (CH₂), 23.8; (CH₃).

Step B: (2-Bromo-3-methyl-5-nitrophenyl)methanamine

4.4 mL of hydrazine hydrate was added to2-[(2-bromo-3-methyl-5-nitrophenyl)methyl]-2,3-dihydro-1H-isoindole-1,3-dione(4.4 g, 11.7 mmol) in 220 mL of acetonitrile. The mixture was stirredunder reflux overnight and then was allowed to cool to room temperature.The phtalhydrazide side product was remove by filtration and washed with100 mL portions of acetonitrile. The combined acetonitrile filtrateswere evaporated and the residue was partitioned between water and ethylacetate. The organic layer was extracted with two 100 mL portions ofaqueous HCl 1 M. The combined acidic aqueous layers were basified topH=10 with solid potassium hydroxide and then extracted with ethylacetate. The organic layer was dried over Na₂SO₄ filtered and evaporatedto yield expected compound as an orange solid (2.15 g, 75% yield). m/z(ESI) 245.0 [M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ8.10; (d, 1H, J=2.8 Hz), 7.94; (d, 1H, J=2.6Hz), 3.96; (s, 2H), 2.47; (s, 3H); ¹³C NMR (75 MHz, CDCl₃) δ146.8; (C),144.4;(C), 140.4;(C), 132.9;(C), 123.4; (CH), 120.6; (CH), 47.0; (CH₂),23.8; (CH₃).

Step C: tert-Butyl N-[(2-bromo-3-methyl-5-nitrophenyl)methyl]carbamate

Di-tert-butyl dicarbonate (1.51 g, 6.90 mmol) was added to(2-bromo-3-methyl-5-nitrophenyl)methanamine (1.54 g, 1.1 eq) in 60 mL ofTHF. The mixture was stirred overnight. After evaporation of thesolvent, the residue was thoroughly washed with pentane and filtered offto yield expected compound as off-white powder (1.93 g, 89% yield). m/z(ESI) 369.0 [M+Na]⁺;

¹H NMR (300 MHz, CDCl₃) δ7.95; (s, 2H), 5.12; (br, 1H, NH), 4.37; (d,2H, J=6.3 Hz), 2.45; (s, 3H), 1.41; (s, 9H); ¹³C NMR (75 MHz, CDCl₃)δ155.7; (C), 146.8; (C), 140.5; (C), 140.4; (C), 132.5; (C), 123.7;(CH), 120.4; (CH), 45.2; (CH₂), 28.3;(OH₃), 23.7; (CH₃).

Step D: tert-ButylN-{[3-methyl-2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methyl}carbamate

In a oven-dried flask and under a nitrogen atmosphere, were placedtert-butyl N-[(2-bromo-3-methyl-5-nitrophenyl)methyl]carbamate (150 mg,0.43 mmol), (+/−) BINAP (14 mg, 0.05 eq), Cs₂CO₃ (196 mg, 1.4 eq),Pd₂dba₃ (10 mg, 0.05 eq) and 3 mL of dry 1,4-dioxanne.N-Methylpiperazine (58 μL, 1.2 eq) was added and the mixture was stirredat 90° C. for 16 h. The solution was filtered through a celite pad andevaporated. The residue was purified by flash chromatography on silicagel (DCM/MeOH/NEt₃//9/1/0.1) to yield expected compound as a pale brownsolid (100 mg, 64% yield). m/z (ESI) 365.2 [M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ7.91; (s, 1H), 7.82; (s, 1H), 5.04; (br, 1H,NH), 4.34; (d, 2H, J=6.1 Hz), 3.26; (br, 2H), 2.90; (br, 2H), 2.61; (br,2H), 2.34-2.29; (m, 8H), 1.40; (s, 9H); ¹³C NMR (75 MHz, CDCl₃) δ156.0;(C), 153.5; (C), 144.6; (C), 139.1; (C), 137.9; (C), 125.6; (CH), 120.5;(CH), 55.8; (CH₂), 49.6; (CH₂), 46.4; (CH₃), 41.5; (CH₂), 28.3; (CH₃),20.1; (CH₃).

Step E: [3-Methyl-2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methanamine

1.2 mL of acetyl chloride was slowly added to 10 mL of MeOH at 0° C. Thesolution was stirred for 30 min at room temperature and then tert-butylN-{[3-methyl-2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methyl}carbamate(300 mg, 0.82 mmol) dissolved in 5 mL of MeOH was added. The reactionmixture was stirred for 2 h and then evaporated. The residue wasdissolved in 15 mL of water and washed with diethyl ether. The aqueouslayer was made alkaline (pH=10) with an aqueous NaOH 1M and extractedwith DCM. The combined organic layers were dried over Na₂SO₄, filteredand evaporated to yield expected compound as a pale yellow oil (216 mg,quantitative yield). m/z (ESI) 265.2 [M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ8.05; (d, J=2.9 Hz, 1H), 7.85; (d, J=2.8 Hz,1H), 3.89; (s, 2H), 3.40-2.80; (m, 4H), 2.70-2.30; (m, 4H), 2.37; (s,3H), 2.32; (s, 3H); ¹³C NMR (75 MHz, CDCl₃) δ153.9; (C), 144.6; (C),143.1; (C), 137.9; (C), 125.2; (CH), 121.3; (CH), 55.9; (CH₂), 49.8;(CH₂), 46.4; (CH₃), 43.4; (CH₂), 20.1; (CH₃).

Step F:1-methyl-4-[2-methyl-4-nitro-6-(pyrrolidin-1-ylmethyl)phenyl]piperazine

A mixture of[3-methyl-2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methanamine (188 mg,0.71 mmol), 1,4-dibromobutane (93 μL, 1.1 eq) and K₂CO₃ (490 mg, 5 eq)in 15 mL of acetonitrile was refluxed for 48 h. After filtration andevaporation of the filtrate, the residue was purified by flashchromatography (DCM/MeOH/NH₄OH//9/1/0.1) to yield expected compound as apale yellow oil (120 mg, 53% yield).

¹H NMR (300 MHz, CDCl₃) δ8.02; (d, J=2.9 Hz, 1H), 7.76; (d, J=2.8 Hz,1H), 3.58; (s, 2H), 3.10-3.00; (m, 4H), 2.35-2.45; (m, 8H), 2.28; (s,3H), 2.25; (s, 3H), 1.70-1.60; (m, 4H); ¹³C NMR (75 MHz, CDCl₃) δ154.5;(C), 143.8; (C), 139.0; (C), 137.3; (C), 125.0; (CH), 123.5; (CH), 57.0;(CH₂), 55.8; (CH₂), 54.0; (CH₂), 49.7; (CH₂), 46.7; (CH₃), 23.5; (CH₂),20.3; (CH₃).

Step G:3-Methyl-4-(4-methylpiperazin-1-yl)-5-(pyrrolidin-1-ylmethyl)aniline

1-Methyl-4-[2-methyl-4-nitro-6-(pyrrolidin-1-ylmethyl)phenyl]piperazine(90 mg, 0.28 mmol) was hydrogenated using ammonium formate (107 mg, 6eq) and Pd/C (10% Pd, 15 mg, 0.05 eq) in 2 mL of EtOH. The mixture wasstirred overnight at room temperature and then filtered through a celitepad. The filtrate was evaporated and the residue was dissolved in DCMand washed with a saturated aqueous solution of Na₂CO₃. The aqueouslayer was extracted with DCM. The combined organic layers were driedover Na₂SO₄, filtered and evaporated to yield expected compound as apale yellow oil (84 mg, quantitative yield).

¹H NMR (300 MHz, CDCl₃) δ6.65; (d, J=2.8 Hz, 1H), 6.30; (d, J=2.8 Hz,1H), 3.64; (s, 2H), 3.45; (br, 2H, NH2), 3.20-3.10; (m, 2H), 2.95-2.85;(m, 2H), 2.58-2.45; (m, 6H), 2.37-2.27; (m, 2H), 2.28; (s, 3H), 2.20;(s, 3H), 1.77-1.67; (m, 4H).

Step H:7-Chloro-N-[3-methyl-4-(4-methylpiperazin-1-yl)-5-(pyrrolidin-1-ylmethyl)phenyl]quinolin-4-amine

3-Methyl-4-(4-methylpiperazin-1-yl)-5-(pyrrolidin-1-ylmethyl)aniline (70mg, 0.24 mmol) and 4,7-dichloroquinoline (50 mg, 1 eq) were refluxedovernight in 5 mL of acetonitrile with 1.25 mL of HCl 1M. The reactionmixture was then evaporated and purified by flash chromatography(DCM/MeOH/NH₄OH//9/1/0.1) to yield expected compound as a white solid(92 mg, 84% yield). m/z (ESI) 450.1 [M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ8.55; (d, J=5.3 Hz, 1H), 8.03; (d, J=2.1 Hz,1H), 7.83; (d, J=9.0 Hz, 1H), 7.45; (dd, J=2.1 and 9.0 Hz, 1H), 7.27;(m, 1H), 7.02; (d, J=2.7 Hz, 1H), 6.96; (d, J=5.3 Hz, 1H), 6.57; (br,1H, NH), 3.76; (s, 2H), 3.30-3.20; (m, 2H), 3.15-3.05; (m, 2H),2.70-2.55; (m, 6H), 2.52-2.42; (m, 2H), 2.40; (s, 3H), 2.38; (s, 3H),1.85-1.75; (m, 4H).

Example 6 Preparation of7-Chloro-N[3-methyl-4-(4-methylpiperazin-1-yl)-5-(morpholin-4-ylmethyl)phenyl]quinolin-4-amine(compound 60) Step A:4-{[3-Methyl-2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methyl}morpholinepiperazine

A mixture of[3-methyl-2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methanamine preparedin step E of example 5 (465 mg, 1.80 mmol), bis(2-chloroethyl) ether(240 μL, 1.2 eq), K₂CO₃ (1.2 g, 5 eq) and Nal (650 mg, 2.3 eq) in 30 mLof acetonitrile was refluxed for 48 h. After filtration and evaporationof the filtrate, the residue was purified by flash chromatography(DCM/MeOH/NH₄O//95/5/1) to yield expected compound as a pale yellow oil(139 mg, 23% yield). m/z (ESI) 335.3 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ8.02; (d, J=2.9 Hz, 1H), 7.76; (d, J=2.8 Hz,1H), 3.58; (s, 2H), 3.10-3.00; (m, 4H), 2.35-2.45; (m, 8H), 2.28; (s,3H), 2.25; (s, 3H), 1.70-1.60; (m, 4H); ¹³C NMR (75 MHz, CDCl₃) δ154.5;(C), 143.8; (C), 139.0; (C), 137.3; (C), 125.0; (CH), 123.5; (CH), 57.0;(CH₂), 55.8; (CH₂), 54.0; (CH₂), 49.7; (CH₂), 46.7; (CH₃), 23.5; (CH₂),20.3; (CH₃).

Step B:3-Methyl-4-(4-methylpiperazin-1-yl)-5-(morpholin-4-ylmethyl)aniline

4-{[3-Methyl-2-(4-methylpiperazin-1-yl)-5-nitrophenyl]methyl}morpholine-piperazine(113 mg, 0.34 mmol) was hydrogenated using ammonium formate (213 mg, 10eq) and Pd/C (10% Pd, 36 mg, 0.05 eq) in 5 mL of EtOH. The mixture wasstirred overnight at room temperature and then filtered through a celitepad. The filtrate was evaporated and the residue was dissolved in DCMand washed with a saturated aqueous solution of Na₂CO₃. The aqueouslayer was extracted with DCM. The combined organic layers were driedover Na₂SO₄, filtered and evaporated to yield expected compound as apale yellow oil (92 mg, 89% yield).

¹H NMR (300 MHz, CDCl₃) δ6.61; (d, J=2.8 Hz, 1H), 6.37; (d, J=2.8 Hz,1H), 3.64-3.75; (m, 4H), 3.49; (s, 3H), 3.16-3.26; (m, 2H), 2.90-3.00;(m, 2H), 2.55-2.65; (m, 2H), 2.30-2.50; (m, 6H), 2.34; (s, 3H), 2.25;(s, 3H); ¹³C NMR (75 MHz, CDCl₃) δ143.7; (C), 140.4; (C), 138.7; (C),138.3; (C), 117.0; (CH), 114.7; (CH), 67.3; (CH₂), 59.8; (CH₂), 56.5;(CH₂), 54.0; (CH₂), 50.3; (CH₂), 46.8; (CH₃), 19.9; (CH₃).

Step C:7-Chloro-N-[3-methyl-4-(4-methylpiperazin-1-yl)-5-(morpholin-4-ylmethyl)phenyl]quinolin-4-amine

3-Methyl-4-(4-methylpiperazin-1-yl)-5-(morpholin-4-ylmethyl)aniline (79mg, 0.26 mmol) and 4,7-dichloroquinoline (56 mg, 1.1 eq) were refluxedovernight in 5 mL of acetonitrile with 0.78 mL of HCl 1M. The reactionmixture was then evaporated and purified by flash chromatography(DCM/MeOH/NH₄OH//9/1/0.1) to yield expected compound as a pale yellowsolid (75 mg, 62% yield). m/z (ESI) 466.4 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.33; (d, J=5.6 Hz, 1H), 8.24; (d, J=9.0 Hz,1H), 7.83; (d, J=2.1 Hz, 1H), 7.46; (dd, J=2.2 and 9.1 Hz, 1H), 7.25;(d, J=2.6 Hz, 1H), 7.06; (d, J=2.5 Hz, 1H), 6.85; (d, J=5.6 Hz, 1H),3.75-3.62; (m, 4H), 3.58; (s, 2H), 3.32-3.20; (m, 2H), 3.17-3.02; (m,2H), 2.75-2.62; (m, 2H), 2.58-2.40; (m, 6H), 2.38; (s, 3H), 2.37; (s,3H); ¹³C NMR (75 MHz, MeOH-d₄) δ151.2; (CH), 150.2; (C), 149.1; (C),145.9; (C), 139.0; (C), 138.2; (C), 136.8; (C), 135.6; (C), 126.7; (CH),125.5; (CH), 125.5; (CH), 123.6; (CH), 123.5; (CH), 118.4; (C), 101.5;(CH), 67.0; (CH₂), 59.8; (CH₂), 56.1; (CH₂), 53.9; (CH₂), 49.5; (CH₂),45.5; (CH₃), 19.0; (CH₃).

Example 77-Chloro-N-[3-methyl-4-(morpholin-4-yl)-5-(pyrrolidin-1-ylmethyl)phenyl]quinolin-4-amine(compound 61) Step A: tert-Butyl N-{[3-methyl-2-(morpholin-4-yl)-5-nitrophenyl]methyl}carbamate

In a oven-dried flask and under a nitrogen atmosphere, were placedtert-butyl N-[(2-bromo-3-methyl-5-nitrophenyl)methyl]carbamate preparedin step C of example 5 (500 mg, 1.45 mmol), (+/−) BINAP (45 mg, 0.07mmol), Cs₂CO₃ (661 mg, 2.03 mmol), Pd₂dba₃ (33 mg, 0.07 eq) and 10 mL ofdry 1,4-dioxanne. Morpholine (152 μL, 1.74 mmol) was added and themixture was stirred at 90° C. for 24 h. The solution was filteredthrough a celite pad and evaporated. The residue was purified by flashchromatography on silica gel (DCM/AcOEt//9/1) to yield expected compoundas a pale brown solid (360 mg, 71% yield); m/z (ESI) 352.2 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ8.00; (d, 1H, J=2.1 Hz), 7.92; (d, 1H, J=2.7Hz), 5.05; (br, 1H, NH), 4.46; (d, 2H, J=6.0 Hz), 3.83; (br, 4H),3.50-2.70; (m, 4H), 2.45; (s, 3H), 1.46; (s, 9H); ¹³C NMR (75 MHz,CDCl₃) δ156.5; (C), 153.5; (C), 145.4; (C), 139.8; (C), 138.6; (C),126.3; (CH), 121.3; (CH), 68.3; (CH₂), 50.6; (CH₂), 42.1; (CH₂), 29.0;(CH₃), 20.7; (CH₃).

Step B: [3-Methyl-2-(morpholin-4-yl)-5-nitrophenyl]methanaminehydrochloride

0.81 mL of acetyl chloride was slowly added to 5 mL of MeOH at 0° C. Thesolution was stirred for 30 min at room temperature and then tert-butylN-{[3-methyl-2-(morpholin-4-yl)-5-nitrophenyl]methyl}carbamate (200 mg,0.57 mmol) dissolved in 5 mL of MeOH was added. The reaction mixture wasstirred for 3 h and then evaporated to yield expected compound as anorange solid (162 mg, 99% yield).

Step C: 4-[2-Methyl-4-nitro-6-(pyrrolidin-1-ylmethyl)phenyl]morpholine

A mixture of [3-Methyl-2-(morpholin-4-yl)-5-nitrophenyl]methanaminehydrochloride (160 mg, 0.56 mmol), 1,4-dibromobutane (80 μL, 0.67 mmol)and K₂CO₃ (542 mg, 3.92 mmol) in 10 mL of acetonitrile was heated at 60°C. for 48 h. After filtration and evaporation of the filtrate, theresidue was purified by flash chromatography (DCM/MeOH/NH₄OH//9/1/0.1)to yield expected compound as a pale yellow oil (100 mg, 58% yield); m/z(ESI) 306.3 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.16; (d, J=3.0 Hz, 1H), 7.98; (d, J=2.7 Hz,1H), 3.90-3.78; (m, 6H), 3.20; (br, 4H), 2.65-2.52; (m, 4H), 2.48; (s,3H), 1.90-1.80; (m, 4H); ¹³C NMR (75 MHz, MeOH-d₄) δ154.5; (C), 144.6;(C), 139.2; (C), 138.4; (C), 125.0; (CH), 123.3; (CH), 67.8; (CH₂),57.0; (CH₂), 54.1; (CH₂), 50.4; (CH₂), 23.4; (CH₂), 19.3; (CH₃).

Step D: 3-Methyl-4-(morpholin-4-yl)-5-(pyrrolidin-1-ylmethyl)aniline

4-[2-Methyl-4-nitro-6-(pyrrolidin-1-ylmethyl)phenyl]morpholine (62 mg,0.20 mmol) was hydrogenated using ammonium formate (128 mg, 2.00 mmol)and Pd/C (10% Pd, 21 mg, 0.02 mmol) in 2 mL of EtOH. The mixture wasstirred overnight at room temperature and then filtered through a celitepad. The filtrate was evaporated and the residue was dissolved in DCMand washed with a saturated aqueous solution of Na₂CO₃. The aqueouslayer was extracted with DCM. The combined organic layers were driedover Na₂SO₄, filtered and evaporated to yield expected compound as apale yellow oil (55 mg, 99% yield); m/z (ESI) 276.3 [M+H]⁺.

Step E:7-Chloro-N-[3-methyl-4-(morpholin-4-yl)-5-(pyrrolidin-1-ylmethyl)phenyl]quinolin-4-amine

3-Methyl-4-(morpholin-4-yl)-5-(pyrrolidin-1-ylmethyl)aniline (46 mg,0.17 mmol) and 4,7-dichloroquinoline (40 mg, 0.20 mmol) were refluxedovernight in 5 mL of acetonitrile with 0.51 mL of HCl 1M. The reactionmixture was then evaporated and purified by flash chromatography(DCM/MeOH/NH₄OH//95/5/0 to 90/10/1) to yield expected compound as awhite solid (53 mg, 71% yield); m/z (ESI) 437.3 [M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ8.43; (d, J=5.5 Hz, 1H), 8.24; (d, J=9.0 Hz,1H), 7.90; (d, J=1.9 Hz, 1H), 7.55; (d, J=2.1 Hz, 1H), 7.30; (dd, J=1.9and 9.0 Hz, 1H), 7.15; (d, J=2.0 Hz, 1H), 6.89; (d, J=5.5 Hz, 1H), 4.15;(s, 1H), 3.89; (d, J=10.9 Hz, 2H), 3.69; (t, J=9.2 Hz, 2H), 3.38; (t,J=9.4 Hz, 2H), 3.01; (br, 4H), 2.74; (d, J=12.0 Hz, 2H), 2.37; (s, 3H),1.98; (br, 4H); ¹³C NMR (75 MHz, CDCl₃) δ151.0; (CH), 148.8; (C), 148.7;(C), 144.3; (C), 138.8; (C), 137.9; (C), 135.8; (C), 134.4; (C), 127.6;(CH), 126.1; (CH), 125.8; (CH), 123.6; (CH), 122.6; (CH), 118.5; (C),102.5; (CH), 68.2; (CH₂), 55.1; (CH₂), 54.3; (CH₂), 50.8; (CH₂), 23.7;(CH₂), 20.5; (CH₃).

Example 8 Preparation of5-[(7-Chloroquinolin-4-yl)amino]-1-N,3-N-bis[3-(4-methylpiperazin-1-yl)propyl]benzene-1,3-dicarboxamide(compound 62) Step A: 1,3-Dimethyl5-[(7-chloroquinolin-4-yl)amino]benzene-1,3-dicarboxylate

1,3-dimethyl 5-aminobenzene-1,3-dicarboxylate (2.09 g, 10 mmol) and4,7-dichloroquinoline (1.98 g, 1 eq) were refluxed in 200 mL of ethanolfor 3 h. The reaction mixture was then cooled to room temperature andthe precipitate was removed by filtration and washed successively with asaturated aqueous solution of NaHCO₃, water, ethanol and then petroleumether to yield expected compound as a yellow powder (2.90 g, 78% yield).m/z (ESI) 371.0 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ8.37 (s, 1H), 8.29; (d, J=9.0 Hz, 1H), 8.07;(d, J=6.9 Hz, 1H), 7.99-7.97; (m, 2H), 7.72-7.70; (m, 1H), 7.46; (d,J=9.3 Hz, 1H), 6.59; (d, J=6.9 Hz, 1H), 3.67; (s, 6H).

Step B:5-[(7-Chloroquinolin-4-yl)amino]-1-N,3-N-bis[3-(4-methylpiperazin-1-yl)propyl]benzene-1,3-dicarboxamide

Synthesized from 1,3-dimethyl5-[(7-chloroquinolin-4-yl)amino]benzene-1,3-dicarboxylate (0.2 g, 0.54mmol) according to general procedure A (reflux for 24 h) with 2 eq ofAlMe₃ and amine in 5 mL of DCM. The residue was purified by preparativethin-layer chromatography (acetone/NH₄OH//9/1) to yield expectedcompound (334 mg, 51%) as a brown oil. m/z(ESI) 621.2 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.31; (d, J=5.5 Hz, 1H), 8.20; (d, J=9.1 Hz,1H), 7.99-7.95; (m, 1H), 7.90-7.84; (m, 2H), 7.74; (d, J=2.0 Hz, 1H),7.37; (dd, J=9.0 and 2.0 Hz, 1H), 6.95; (d, J=5.5 Hz, 1H), 3.45-3.30;(m, 4H), 2.70-2.21; (m, 20H), 2.15; (s, 6H), 1.80-1.60; (m, 4H); ¹³C NMR(75 MHz, MeOH-d₄) δ167.6; (C), 151.8; (CH), 149.3; (C), 149.1; (C),141.4; (C), 136.8; (C), 136.0; (CH), 127.1; (CH), 126.2; (CH), 124.1;(CH), 124.0; (CH), 121.8; (CH), 119.0; (C), 102.7; (CH), 56.4; (CH₂),54.7; (CH₂), 52.7; (CH₂), 45.0; (CH₃), 38.9; (CH₂), 26.2; (CH₂).

Another fraction give methyl3-[(7-chloroquinolin-4-yl)amino]-5-[(4-methylpiperazin-1-yl)carbonyl]benzoate(example 67) (31 mg, 12%) as a yellow powder.

Example 9 Preparation ofN-{3,5-Bis[(4-methylpiperazin-1-yl)carbonyl]phenyl}-7-chloroquinolin-4-amine(compound 63)

Synthesized from 1,3-dimethyl5-[(7-chloroquinolin-4-yl)amino]benzene-1,3-dicarboxylate prepared fromstep A of example 8 (0.2 g, 0.54 mmol) according to general procedure A(reflux for 24 h) with 1 eq of AlMe₃ and amine in 5 mL of DCM. Theresidue was purified by preparative thin-layer chromatography(acetone/NH₄OH//9/1) to yield expected compound (273 mg, 55%) as ayellow oil. m/z(ESI) 507.0 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.53; (d, J=5.5 Hz, 1H), 8.31; (d, J=9.1 Hz,1H), 7.95; (d, J=2.1 Hz, 1H), 7.58; (dd, J=9.1 and 2.2 Hz, 1H), 7.52;(d, J=1.4 Hz, 2H), 7.25; (t, J=1.4 Hz, 1H), 7.20; (d, J=5.5 Hz, 1H),3.81; (br, 4H), 3.57; (br, 4H), 2.63-2.40; (m, 8H), 2.36; (s, 6H).

Another fraction give methyl3-[(7-chloroquinolin-4-yl)amino]-5-[(4-methylpiperazin-1-yl)carbonyl]benzoate(compound 68) (235 mg, 55%) as a yellow oil.

Example 10 Preparation ofN-[3,5-Bis({4-[3-(dimethylamino)propyl]piperazin-1-yl}carbonyl)phenyl]-7-chloroquinolin-4-amine(compound 64)

Synthesized from 1,3-dimethyl5-[(7-chloroquinolin-4-yl)amino]benzene-1,3-dicarboxylate prepared fromstep A of example 8 (0.3 g, 0.81 mmol) according to general procedure A(reflux for 48 h) with 2.5 eq of AlMe₃ and amine in 10 mL of DCM. Theresidue was purified by preparative thin-layer chromatography(acetone/NH₄OH//9/1) to yield expected compound (12 mg, 2%) as a yellowoil. m/z(MALDI-TOF) 649.4 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.52; (d, J=5.5 Hz, 1H), 8.30; (d, J=9.1 Hz,1H), 7.93; (d, J=2.1 Hz, 1H), 7.57; (dd, J=9.1 and 2.1 Hz, 1H), 7.51;(d, J=1.3 Hz, 2H), 7.23; (t, J=1.3 Hz, 1H), 7.19; (d, J=5.5 Hz, 1H),3.81; (br, 4H), 3.56; (br, 4H), 2.65-2.35; (m, 16H), 2.28; (s, 12H),1.80-1.65; (m, 4H); ¹³C NMR (75 MHz, MeOH-d₄) δ169.7; (C), 151.7; (CH),149.7; (C), 148.9; (C), 141.8; (C), 137.7; (C), 136.1; (C), 127.1; (CH),126.3; (CH), 123.8; (CH), 121.6; (CH), 120.6; (CH), 119.2; (C), 103.2;(CH), 57.5; (CH₂), 56.2; (CH₂), 48.0; (CH₂), 44.4; (CH₃), 42.2; (CH₂),24.3; (CH₂).

Another fraction give methyl3-[(7-chloroquinolin-4-yl)amino]-5-({4-[3-(dimethylamino)propyl]piperazin-1-yl}carbonyl)benzoate(compound 69) (53 mg, 13%) as a yellow oil.

Example 11 Preparation ofN-[3,5-Bis(pyrrolidin-1-ylmethyl)phenyl]-7-chloroquinolin-4-amine(compound 65) Step A: 3,5-Bis[(pyrrolidin-1-yl)carbonyl]aniline

A solution of AlMe₃ (2M in toluene, 19.5 mL, 2 eq) was added dropwise toa solution of pyrrolidine (3.25 mL, 2 eq) in 50 mL of anhydrous DCM at0° C. The reaction mixture was allowed to warm up to room temperatureand stirred for 1 h. 1,3-Dimethyl 5-aminobenzene-1,3-dicarboxylate (4.10g, 19.5 mmol) was added and the reaction mixture was refluxed for 38 h.The mixture was then poured in ice cooled water and filtered. Theaqueous filtrate was extracted with CHCl₃. The combined organic layerswere dried over MgSO₄, filtered and evaporated to give 3.5 g of expectedcompound which was used in the next step without further purification.

Step B:N-{3,5-Bis[(pyrrolidin-1-yl)carbonyl]phenyl}-7-chloroquinolin-4-amine

3,5-Bis[(pyrrolidin-1-yl)carbonyl]aniline (287 mg, 1.0 mmol) and4,7-dichloroquinoline (198 mg, 1 eq) were refluxed in 20 mL of ethanoland 2 mL of HCl 1M for 12 h. The reaction mixture was then evaporated.The residue was purified by flash chromatography (DCM/MeOH//9/1) toyield expected compound as a yellow powder (2.90 g, 78% yield). m/z(ESI) 449.1 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ11.50; (br, 1H), 9.08; (d, J=9.0 Hz, 1H),8.30-8.10; (m, 2H), 7.85-7.75; (m, 2H), 7.68-7.60; (m, 1H), 7.44; (d,J=8.4 Hz, 1H), 6.93-6.82; (m, 1H), 3.70-3.48; (m, 8H), 2.03-1.85; (m,8H); ¹³C NMR (75 MHz, CDCl₃) δ167.7; (C), 155.2; (C), 142.3; (CH),140.5; (C), 139.1; (C), 138.7; (C), 137.6; (C), 128.3; (CH), 126.8;(CH), 125.3; (CH), 124.8; (CH), 119.6; (CH), 116.4; (C), 100.5; (CH),50.0; (CH₂), 46.9; (CH₂), 26.6; (CH₂), 24.5; (CH₂).

Step C:N-[3,5-Bis(pyrrolidin-1-ylmethyl)phenyl]-7-chloroquinolin-4-amine

To a suspension ofN-{3,5-bis[(pyrrolidin-1-yl)carbonyl]phenyl}-7-chloroquinolin-4-amine(84 mg, 0.19 mmol) in 15 mL of anhydrous THF, was added portionwiseLiAlH₄ (71 mg, 10 eq). The reaction mixture was stirred at roomtemperature for 2 h and then 10 mL of ethyl acetate was added. Themixture was filtered through a celite pad. The filtrate was evaporatedand the residue was purified by preparative thin-layer chromatography(DCM/MeOH/NH₄OH//95/5/1) to yield expected compound (4 mg, 5%) as ayellow oil. m/z(ESI) 420.9 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ8.56; (d, J=5.3 Hz, 1H), 8.01; (d, J=2.1 Hz,1H), 7.89; (d, J=9.0 Hz, 1H), 7.44; (dd, J=8.9 and 2.1 Hz, 1H),7.30-7.22; (m, 2H), 7.14-7.10; (m, 1H), 7.03-6.80; (m, 2H), 2.70-2.55;(m, 8H), 1.90-1.78; (m, 8H).

Example 12 Preparation of 4-[(7-Chloroquinolin-4-yl)amino]-2,6-bis(pyrrolidin-1-ylmethyl)phenol (compound 66) Step A:4-[(7-Chloroquinolin-4-yl)amino]phenol

4-Aminophenol (1.09 g, 10 mmol) and 4,7-dichloroquinoline (1.98 g, 1 eq)were refluxed in 50 mL of ethanol for 2 h. The reaction mixture was thencooled to room temperature and the precipitate was removed by filtrationand washed successively with a saturated aqueous solution of NaHCO₃,water, methanol and then petroleum ether to yield expected compound as ayellow powder (2.57 g, 95% yield). m/z (ESI) 271.1 [M+H]⁺.

¹H NMR (300 MHz, DMSO-d₆) δ10.7; (br, 1H), 9.88; (br, 1H), 8.77; (d,J=9.1 Hz, 1H), 8.43; (d, J=6.8 Hz, 1H), 8.09; (d, J=2.1 Hz, 1H), 7.77(dd, J=9.1 and 2.1 Hz, 1H), 7.23; (d, J=8.8 Hz, 1H), 6.94; (d, J=8.8 Hz,1H), 6.60; (d, J=6.8 Hz, 1H).

Step B:4-[(7-Chloroquinolin-4-yl)amino]-2,6-bis(pyrrolidin-1-ylmethyl)phenol

4-[(7-Chloroquinolin-4-yl)amino]phenol (2.45 g, 9 mmol), pyrrolidine(3.25 mL, 4.4 eq) and a 37% aqueous solution of formaldehyde (3 mL, 4.4eq) were stirred at room temperature for 18 h in 5 mL of ethanol. Thereaction mixture was then evaporated and the residue was purified byflash chromatography (DCM/MeOH/NH₄OH/8:2:0.1) to yield expected compoundas a brown solid (2.90 g, 74% yield). m/z (ESI) 437.1 [M+H]⁺.

¹H NMR (300 MHz, DMSO-d₆) δ8.91; (br, 1H), 8.41; (d, J=9.0 Hz, 1H),8.36; (d, J=5.4 Hz, 1H), 7.84; (d, J=2.1 Hz, 1H), 7.51; (dd, J=9.0 and2.1 Hz, 1H), 7.06; (s, 2H), 6.60; (d, J=5.4 Hz, 1H), 2.68-2.52; (m, 8H),1.83-1.60; (m, 8H).

Example 13 Preparation of Methyl3-[(7-chloroquinolin-4-yl)amino]-5-[(4-methylpiperazin-1-yl)carbonyl]benzoate(compound 67)

Prepared as described in step B of example 8.

m/z(ESI) 496.0 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.47; (d, J=5.5 Hz, 1H), 8.30; (d, J=9.1 Hz,1H), 8.26-8.20; (m, 1H), 8.18-8.12; (m, 1H), 8.10-8.03; (m, 1H), 7.90;(d, J=1.4 Hz, 1H), 7.54; (dd, J=9.0 and 1.4 Hz, 1H), 7.08; (d, J=5.5 Hz,1H), 3.96; (s, 3H), 3.46; (t, J=6.8 Hz, 2H), 2.60-2.40; (m, 10H), 2.27;(s, 3H), 1.84; (q, J=7.1 Hz, 2H); ¹³C NMR (75 MHz, MeOH-d₄) δ167.4; (C),166.3; (C), 151.5; (CH), 149.1; (C), 149.0; (C), 141.5; (C), 136.6; (C),136.0; (C), 132.1; (C), 126.8; (CH), 126.2; (CH), 125.8; (CH), 125.5;(CH), 124.0; (CH), 123.5; (CH), 118.9; (C), 102.5; (CH), 55.9; (CH₂),54.2; (CH₂), 52.1; (CH₃), 51.9; (CH₂), 44.2; (CH₃), 38.6; (CH₂), 25.9;(CH₂).

Example 14 Preparation of Methyl3-[(7-chloroquinolin-4-yl)amino]-5-[(4-methylpiperazin-1-yl)carbonyl]benzoate(compound 68)

Prepared as described in example 9.

m/z(ESI) 439.0 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.53; (d, J=5.5 Hz, 1H), 8.32; (d, J=9.1 Hz,1H), 8.14-8.12; (m, 1H), 7.95; (d, J=2.0 Hz, 1H), 7.84-7.82; (m, 1H),7.67-7.65; (m, 1H), 7.58; (dd, J=9.1 and 2.2 Hz, 1H), 7.15; (d, J=5.5Hz, 1H), 3.97; (s, 3H), 3.83; (br, 2H), 3.57; (br, 2H), 2.62-2.40; (m,4H), 2.37; (s, 3H).

Example 15 Preparation of Methyl3-[(7-chloroquinolin-4-yl)amino]-5-({4-[3-(dimethylamino)propyl]piperazin-1-yl}carbonypbenzoate(compound 69)

Prepared as described in example 10.

m/z(MALDI-TOF) 510.3 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.45; (d, J=5.5 Hz, 1H), 8.24; (d, J=9.1 Hz,1H), 8.06-8.03; (m, 1H), 7.88; (d, J=2.1 Hz, 1H), 7.78-7.75; (m, 1H),7.62-7.59; (m, 1H), 7.50; (dd, J=9.1 and 2.2 Hz, 1H), 7.08; (d, J=5.5Hz, 1H), 3.93; (s, 3H), 3.79; (br, 2H), 3.52; (br, 2H), 2.60-2.30; (m,8H), 2.24; (s, 6H), 1.78-1.62; (m, 2H).

Example 16 Preparation ofN[3,5-bis(4-methylpiperazin-1-yl)phenyl]-7-chloroquinolin-4-amine(compound 70) Step A: 2,6-Dibromo-4-nitroaniline

2,6-Dibromo-4-nitroaniline was prepared from p-nitroaniline as describedby Shepherd, R. G. J. Org. Chem. 1947, 12, 275.

Step B: 3,5-Dibromonitrobenzene

NaNO₂ (4.1 g, 59.1 mmol) was added portionwise to a solution of2,6-dibromo-4-nitroaniline (5 g, 3.5 eq) in 20% H₂SO₄ and AcOEt (each 40mL) at a rate to keep the internal temperature at 50-55° C. Afterheating for an additional 10 min, the mixture was cooled, diluted, andextracted with AcOEt. The combined organic layers were dried overNa₂SO₄, filtered and evaporated. The residue was purified by flashchromatography (cyclohexane/DCM//7/3 then 8/2) to yield expectedcompound as a white solid (1.94 g, 41% yield).

¹H NMR (300 MHz, CDCl₃) δ8.32; (d, J=1.8 Hz, 2H), 8.00; (t, J=1.8 Hz,1H).

Step C: 1-methyl-4-[3-(4-methylpiperazin-1-yl)-5-nitrophenyl]piperazine

In a oven-dried flask and under a nitrogen atmosphere, were placed3,5-dibromonitrobenzene (500 mg, 1.78 mmol), (+/−) BINAP (110 mg, 0.1eq), Cs₂CO₃ (1.62 g, 3 eq), Pd₂dba₃ (81 mg, 0.11 eq) and 10 mL of dry1,4-dioxane. N-Methylpiperazine (0.47 mL, 2.4 eq) was added and themixture was stirred at 90° C. for 48 h. The solution was filteredthrough a celite pad and evaporated. The residue was purified by flashchromatography on silica gel (DCM/MeOH/NH₄OH//95/5/1) to yield expectedcompound as an orange solid (258 mg, 45% yield). m/z (MALDI-TOF) 320.2[M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ7.17 (d, J=2.1 Hz, 2H), 6.59 (t, J=2.1 Hz, 1H),3.20; (t, J=4.8 Hz, 8H), 2.51; (t, J=5.1 Hz, 8H), 2.29; (s, 6H); ¹³C NMR(75 MHz, CDCl₃) δ152.4; (C), 150.1; (C), 108.1; (CH), 101.8; (CH), 54.8;(CH₂), 48.7; (CH₂), 46.1; (CH₃).

Step D: 3,5-bis(4-methylpiperazin-1-yl)aniline

1-methyl-4-[3-(4-methylpiperazin-1-yl)-5-nitrophenyl]piperazine (258 mg,0.81 mmol) was hydrogenated using ammonium formate (0.51 g, 10 eq) andPd/C (10% Pd, 85 mg, 0.1 eq) in 8 mL of EtOH. The mixture was stirredovernight at room temperature and then filtered through a celite pad.The filtrate was evaporated and the residue was dissolved in DCM andwashed with a saturated aqueous solution of Na₂CO₃. The aqueous layerwas extracted with DCM. The combined organic layers were dried overNa₂SO₄, filtered and evaporated to yield expected compound as a brownpowder (212 mg, 90% yield). m/z (MALDI-TOF) 290.2 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ5.97; (t, J=1.8 Hz, 1H), 5.84; (d, J=2.1 Hz,2H), 3.17; (t, J=4.8 Hz, 8H), 2.54; (t, J=5.1 Hz, 8H), 2.34; (s, 6H);¹³C NMR (75 MHz, CDCl₃) δ153.6; (C), 148.1; (C), 96.4; (CH), 96.2; (CH),(CH₂ (CH₂), 46.5; (CH₃).

Step E:N-[3,5-bis(4-methylpiperazin-1-yl)phenyl]-7-chloroquinolin-4-amine

In a oven-dried flask and under a nitrogen atmosphere, were placed3,5-bis(4-methylpiperazin-1-yl)aniline (148 mg, 0.51 mmol),4,7-dichloroquinoline (101 mg, 1 eq), (+/−) BINAP (16 mg, 0.05 eq),Cs₂CO₃ (232 mg, 1.4 eq), Pd₂dba₃ (12 mg, 0.02 eq) and 2 mL of dry1,4-dioxane. The mixture was stirred at 90° C. for 48 hours. Thesolution was filtered through a celite pad and evaporated. The residuewas purified by flash chromatography on silica gel(DCM/MeOH/NH₄OH//8/2/0 then 8/2/0.1). 221 mg of an yellow solid wasobtained. A subsequent purification by preparative thin-layerchromatography (DCM/MeOH/NH₄OH//8/2/0.1) yield expected compound as ayellow solid (65 mg, 28% yield). m/z (ESI) 451.2 [M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ8.53; (d, J=5.4 Hz, 1 H), 8.03; (d, J=2.1 Hz,1H), 7.84; (d, J=8.7 Hz, 1H), 7.44; (dd, J=9.0 and 2.1 Hz, 1H), 6.99;(d, J=5.4 Hz, 1H), 6.60; (br, 1H), 6.35-6.40; (m, 2H), 6.30-6.35; (m,1H), 3.24; (t, J=5.1 Hz, 8H), 2.60; (t, J=5.1 Hz, 8H), 2.37; (s, 6H);¹³C NMR (75 MHz, CDCl₃) δ153.5; (C), 151.7; (CH), 149.4; (C), 148.8;(C), 141.0; (C), 135.6; (C), 128.5; (CH), 126.1; (CH), 122.1; (CH),118.3; (C), 103.0; (CH), 101.2; (CH), 55.3; (CH₂), 49.3; (CH₂), 46.3;(CH₃).

Example 17

Preparation ofN-[3,5-bis(morpholin-4-yl)phenyl]-7-chloro-quinolin-4-amine (compound71) Step A: 4-[3-(morpholin-4-yl)-5-nitrophenyl]morpholine

In a oven-dried flask and under a nitrogen atmosphere, were placed3,5-dibromonitrobenzene prepared from step B of example 16 (150 mg, 0.53mmol), (+/−) BINAP (33 mg, 0.1 eq), Cs₂CO₃ (0.49 g, 3 eq), Pd₂dba₃ (24mg, 0.05eq) and 3 mL of dry 1,4-dioxane. Morpholine (0.11 mL, 2.4eq) wasadded and the mixture was stirred at 90° C. for 5 days. The solution wasfiltered through a celite pad and evaporated. The residue was purifiedby flash chromatography on silica gel (DCM/MeOH/NH₄OH//98/2/1) to yieldexpected compound as an orange solid (123 mg, 51% yield). m/z(MALDI-TOF) 294.1 [M+H]⁺;

¹H NMR (300 MHz, CDCl₃) δ7.26; (s, 2H), 6.65; (s, 1H), 3.87; (t, J=4.8Hz, 8H), 3.22; (t, J=4.8 Hz, 8H).

Step B: 3,5-bis(morpholin-4-yl)aniline

4-[3-(morpholin-4-yl)-5-nitrophenyl]morpholine (123 mg, 0.42 mmol) washydrogenated using ammonium formate (0.26 g, 10 eq) and Pd/C (10% Pd, 45mg, 0.1 eq) in 4 mL of EtOH. The mixture was stirred overnight at roomtemperature and then filtered through a celite pad. The filtrate wasevaporated and the residue was dissolved in DCM and washed with asaturated aqueous solution of Na₂CO₃. The aqueous layer was extractedwith DCM. The combined organic layers were dried over Na₂SO₄, filteredand evaporated to yield expected compound as a brown powder (63 mg, 57%yield).

¹H NMR (300 MHz, CDCl₃) δ5.93; (t, J=1.8 Hz, 1H), 5.84; (d, J=1.8 Hz,2H), 3.83; (t, J=5.1 Hz, 8H), 3.11; (t, J=5.1 Hz, 8H).

Step C: N-[3,5-bis(morpholin-4-yl)phenyl]-7-chloroquinolin-4-amine

In a oven-dried flask and under a nitrogen atmosphere, were placed3,5-bis(morpholin-4-yl)aniline (63 mg, 0.22 mmol), 4,7-dichloroquinoline(43 mg, 1 eq), (+/−) BINAP (7 mg, 0.1 eq), Cs₂CO₃ (99 mg, 1.4 eq),Pd₂dba₃ (5 mg, 0.02 eq) and 2 mL of dry 1,4-dioxane. The mixture wasstirred at 90° C. for 24 h. The solution was filtered through a celitepad and evaporated. The residue was purified by preparative thin-layerchromatography (DCM/MeOH//95/5) to yield expected compound as a yellowsolid (27 mg, 29% yield). m/z (ESI) 425.3 [M+H]⁺;

¹H NMR (300 MHz, MeOH-d₄) δ8.35; (d, J=5.7 Hz, 1H), 8.27; (d, J=9.1 Hz,1H), 7.85; (d, J=2.1 Hz, 1H), 7.49; (dd, J=9.1 and 2.1 Hz, 1H), 6.94;(d, J=5.7 Hz, 1H), 6.47-6.52; (m, 2H), 6.40-6.47; (m, 1H), 3.82; (t,J=4.7 Hz, 8H), 3.15; (t, J=4.9 Hz, 8H); ¹³C NMR (75 MHz, MeOH-d₄)δ153.9; (C), 150.9; (C), 150.7; (CH), 148.4; (C), 141.1; (C), 136.0;(C), 126.1; (CH), 125.8; (CH), 123.8; (CH), 118.3; (C), 103.7; (CH),102.0; (CH), 101.1; (CH), 67.0; (CH₂), 49.7; (CH₂).

Example 18 Preparation of3-[(7-chloroquinolin-4-yl)amino]-5-(4-methyl-piperazin-1-yl)phenol(compound 72)

3,5-bis(4-methylpiperazin-1-yl)aniline prepared from step D of example16 (95 mg, 0.33 mmol) and 4,7-dichloroquinoline (65 mg, 1 eq) wererefluxed 3h in 4 mL of acetonitrile with 0.98 mL of HCl 1 M. Thereaction mixture was then evaporated and purified by flashchromatography (DCM/MeOH/NH₄OH//9/1/0.1) to yield expected compound as ayellow oil (28 mg, 23% yield). m/z (ESI) 369.2 [M+H]⁺;

¹H NMR (300 MHz, MeOH-d₄) δ8.32; (d, J=5.6 Hz, 1H), 8.20; (d, J=9.0 Hz,1H), 7.80; (d, J=2.0 Hz, 1H), 7.42; (dd, J=2.1 and 9.0 Hz, 1H), 6.94;(d, J=5.6 Hz, 1H), 6.40-6.43; (m, 1H), 6.32-6.35; (m, 1H), 6.23-6.26;(m, 1H), 3.15; (t, J=5.1 Hz, 4H), 2.55; (t, J=5.1 Hz, 4H), 2.30; (s,3H).

Example 19 Preparation of3-Amino-5-[(7-chloroquinolin-4-yl)amino]benzonitrile hydrochloride(compound 73) Step A: 3,5-Diaminobenzonitrile

To 3,5-dinitrobenzonitrile (5.0 g, 25.89 mmol) in 20 mL of HCl 1M, wasadded SnCl₂ (34.4 g, 7 eq). The reaction mixture was stirred at roomtemperature for 2 h and then cooled to 0° C. The mixture was madealkaline with a 50% aqueous solution of NaOH and the precipitate wasremoved by filtration. The filtrate was extracted with ethyl acetate andthe combined organic layers were evaporated. The residue and theprecipitate were purified by flash chromatography on silica gel(DCM/MeOH//95/5) to yield expected compound as an orange solid (1.95 g,57% yield). m/z (ESI) 134.0 [M+H]⁺.

Step B: 3-Amino-5-[(7-chloroquinolin-4-yl)amino]benzonitrilehydrochloride

3,5-Diaminobenzonitrile (500 mg, 3.76 mmol) and 4,7-dichloroquinoline(744 mg, 1 eq) were stirred at room temperature in 50 mL of ethanol for18 h. The precipitate was removed by filtration to yield expectedcompound as a yellow powder (925 mg, 74% yield).

¹H NMR (300 MHz, DMSO-d₆) δ11.08; (br, 1H), 8.83; (d, J=9.2 Hz, 1H),8.56; (d, J=7.0 Hz, 1H), 8.17; (d, J=2.0 Hz, 1H), 7.87; (dd, J=9.2 and2.0 Hz, 1H), 7.02-6.98; (m, 1H), 6.95-6.85; (m, 3H), 6.00; (br, 2H); ¹³CNMR (75 MHz, DMSO-d₆) δ158.2; (C), 154.7; (C), 147.1; (CH), 142.7; (C),142.3; (C), 142.0; (C), 131.0; (CH), 129.7; (CH), 122.9; (CH), 122.2;(C), 119.6; (C), 118.5; (CH), 118.2; (CH), 117.8; (CH), 116.3; (C),104.5; (CH).

Example 20 Preparation of4-[(3-amino-5-cyanophenyl)amino]-7-chloro-1-[2-(piperidin-1-yl)ethyl]quinolin-1-iumchloride (compound 74)

3-Amino-5-[(7-chloroquinolin-4-yl)amino]benzonitrile hydrochlorideprepared from step B of example 19 (0.88 g, 2.98 mmol),1-(2-chloroethyl)piperidine hydrochloride (0.55 g, 1 eq) and K₂CO₃ (0.83g, 2 eq) were refluxed in 50 mL of acetonitrile for 48 h. The reactionmixture was then filtered. The filtrate was evaporated to yield expectedcompound as a yellow powder (0.58 g, 44% yield). m/z (ESI) 406.2 [M]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.62; (d, J=9.1 Hz, 1H), 8.46; (d, J=7.4 Hz,1H), 8.39; (d, J=1.9 Hz, 1H), 7.88; (dd, J=9.1 and 1.9 Hz, 1H),7.10-6.90; (m, 3H), 6.94; (d, J=7.4 Hz, 1H), 4.72; (t, J=6.0 Hz, 2H),2.83; (t, J=6.0 Hz, 2H), 2.55-2.40; (m, 4H), 1.62-1.40; (m, 6H).

Example 21 Preparation of 3-amino-5-({7-chloro-1-[2-(piperidin-1-yl)ethyl]-1,4-dihydroquinolin-4 ylidene}amino)benzonitrile (compound 75)

3-Amino-5-[(7-chloroquinolin-4-yl)amino]benzonitrile hydrochlorideprepared from step B of example 19 (200 mg, 0.68 mmol),1-(2-chloroethyl)piperidine hydrochloride (125 mg, 1 eq), K₂CO₃ (468 mg,5 eq) and DIEA (0.11 mL, 1 eq) were refluxed in 30 mL of acetonitrilefor 72 h. The reaction mixture was then filtered. The filtrate wasevaporated and the residue was purified by flash chromatography onsilica gel (AcOEt/MeOH//100/0 to 95/5) to yield expected compound as ayellow powder (8 mg, 3% yield). m/z (ESI) 406.2 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.37; (d, J=8.8 Hz, 1H), 7.63; (d, J=1.9 Hz,1H), 7.40-7.28; (m, 2H), 6.70-6.66; (m, 1H), 6.53-6.50; (m, 1H),6.49-6.46; (m, 1H), 5.88 (d, J=7.9 Hz, 1H), 4.16; (t, J=6.9 Hz, 2H),2.64; (t, J=6.9 Hz, 2H), 2.52-2.40; (m, 4H), 1.64-1.40; (m, 6H).

Example 22 Preparation of 3-({7-Chloro-1-[2-(piperidin-1-yl)ethyl]-1,4-dihydroquinolin-4-ylidene}amino)-5-{[2-(piperidin-1-yl)ethyl]amino}benzonitrile(compound 76)

3-amino-5-({7-chloro-1-[2-(piperidin-1-yl)ethyl]-1,4-dihydroquinolin-4-ylidene}amino)benzonitrileprepared from example 21 (132 mg, 0.32 mmol),1-(2-chloroethyl)piperidine hydrochloride (182 mg, 3 eq) and DIEA (0.51mL, 9.3 eq) were refluxed in 15 mL of 2-methylbutan-1-ol for 10 h. Thereaction mixture was then evaporated and the residue and was purified bypreparative thin-layer chromatography (DCM/MeOH/NH₄OH//95/5/1) to yieldexpected compound as a yellow oil (28 mg, 17% yield). m/z (ESI) 517.5[M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.37; (d, J=8.8 Hz, 1H), 7.64; (d, J=1.7 Hz,1H), 7.36; (d, J=8.0 Hz, 1H), 7.32; (dd, J=8.2 and 1.8 Hz, 1H),6.68-6.59; (m, 1H), 6.50-6.40; (m, 2H), 5.91; (d, J=7.9 Hz, 1H), 4.17;(t, J=6.8 Hz, 2H), 3.25; (t, J=6.8 Hz, 2H), 2.73-2.35; (m, 12H),1.70-1.40; (m, 12H); ¹³C NMR (75 MHz, MeOH⁻d₄) δ156.6; (C), 153.9; (C),151.0; (C), 142.1; (CH), 140.3; (C), 137.8; (C), 127.5; (CH), 124.1;(CH), 123.0; (C), 120.0; (C), 115.5; (CH), 113.5; (CH), 113.4; (C),110.4; (CH), 110.0; (CH), 100.8; (CH), 57.7; (CH₂), 56.7; (CH₂), 54.8;(CH₂), 54.6; (CH₂), 49.8; (CH₂), 40.2; (CH₂), 25.9; (CH₂), 25.6; (CH₂),24.2; (CH₂), 24.1; (CH₂).

Example 23 Preparation of3-[(7-chloroquinolin-4-yl)amino]-5-{[2-(piperidin-1-yl)ethyl]amino}benzonitrile(compound 77) Step A:3-amino-5-{[2-(piperidin-1-yl)ethyl]amino}benzonitrile

3,5-Diaminobenzonitrile prepared from step A of example 19 (511 mg, 3.84mmol), 1-(2-chloroethyl)piperidine hydrochloride (707 mg, 1 eq) and DIEA(1.97 mL, 3 eq) were refluxed in 50 mL of 2-methylbutan-1-ol for 24 h.The reaction mixture was then evaporated to give 751 mg of expectedcompound which was used in the next step without further purification.m/z (ESI) 245.3 [M+H]⁺.

Step B:3-[(7-chloroquinolin-4-yl)amino]-5-{[2-(piperidin-1-yl)ethyl]amino}benzonitrile

3-amino-5-{[2-(piperidin-1-yl)ethyl]amino}benzonitrile (200 mg, 0.82mmol) and 4,7-dichloroquinoline (162 mg, 1 eq) were refluxed in 15 mL ofethanol and 0.82 mL of HCl 1M for 24 h. The reaction mixture was thenevaporated and the residue and was purified by preparative thin-layerchromatography (DCM/MeOH/NH₄OH//95/5/1) to yield expected compound as ayellow powder (51 mg, 15% yield). m/z (ESI) 406.3 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ8.40; (d, J=5.4 Hz, 1H), 7.92; (d, J=9.1 Hz,1H), 7.83; (d, J=2.1 Hz, 1H), 7.24; (dd, J=9.0 and 2.1 Hz, 1H), 6.89;(d, J=5.4 Hz, 1H), 6.72-6.69; (m, 1H), 6.68-6.63; (m, 1H), 6.50-6.43;(m, 1H), 4.97; (br, 1H), 4.60; (br, 1H), 3.10-2.97; (m, 2H), 2.57-2.47;(m, 2H), 2.43-2.22; (m, 4H), 1.57-1.43; (m, 4H), 1.41-1.30; (m, 2H); ¹³CNMR (75 MHz, CDCl₃) δ151.3; (CH), 149.7; (C), 149.1; (C), 147.3; (C),141.7; (C), 135.5; (C), 128.1; (CH), 126.2; (CH), 122.1; (CH), 119.0;(C), 118.4; (C), 113.5; (C), 113.2; (CH), 111.1; (CH), 109.8; (CH),103.4; (CH), 56.2; (CH₂), 53.9; (CH₂), 39.2; (CH₂), 24.9; (CH₂), 23.6;(CH₂).

Example 24 Preparation of4-[(7-chloroquinolin-4-yl)amino]-2,6-bis(pyrrolidin-1-ylmethyl)phenol(compound 78)

Step A: 4-[(7-Chloroquinolin-4-yl)amino]phenol

4-aminophenol (1.09 g, 10 mmol) and 4,7-dichloroquinoline (1.98 g, 1 eq)were refluxed in 50 mL of ethanol for 2 h. The reaction mixture was thencooled to room temperature and the precipitate was removed by filtrationand washed successively with a saturated aqueous solution of NaHCO₃,water, methanol and then petroleum ether to yield expected compound as ayellow powder (2.57 g, 95% yield). LC-MS: m/z (ESI) 271.1 [M+H]⁺.

¹H NMR (300 MHz, DMSO-d₆) δ10.7; (br, 1H), 9.88; (br, 1H), 8.77; (d,J=9.1 Hz, 1H), 8.43; (d, J=6.8 Hz, 1H), 8.09; (d, J=2.1 Hz, 1H), 7.77;(dd, J=9.1 and 2.1 Hz, 1H), 7.23; (d, J=8.8 Hz, 1H), 6.94; (d, J=8.8 Hz,1H), 6.60; (d, J=6.8 Hz, 1H).

Step B:4-[(7-Chloroquinolin-4-yl)amino]-2,6-bis(pyrrolidin-1-ylmethyl)phenol

4-[(7-chloroquinolin-4-yl)amino]phenol (2.45 g, 9 mmol), pyrrolidine(3.25 mL, 4.4 eq) and a 37% aqueous solution of formaldehyde (3 mL, 4.4eq) were stirred at room temperature for 18 h in 5 mL of ethanol. Thereaction mixture was then evaporated and the residue was purified byflash chromatography (DCM/MeOH/NH₄OH 8:2:0.1) to yield expected compoundas a brown solid (2.90 g, 74% yield). LC-MS: m/z (ESI) 437.1 [M+H]⁺.

¹H NMR (300 MHz, DMSO-d₆) δ8.91; (br, 1H), 8.41; (d, J=9.0 Hz, 1H),8.36; (d, J=5.4 Hz, 1H), 7.84; (d, J=2.1 Hz, 1H), 7.51; (dd, J=9.0 and2.1 Hz, 1H), 7.06; (s, 2H), 6.60; (d, J=5.4 Hz, 1H), 2.68-2.52; (m, 8H),1.83-1.60; (m, 8H).

Example 25 Preparation of4-[(7-chloroquinolin-4-yl)amino]-2,6-bis(morpholin-1-ylmethyl)phenol(compound 79)

4-[(7-Chloroquinolin-4-yl)amino]phenol (of example 24—step A) (0.292 g,1.08 mmol), morpholine (0.89 mL, 8.8 eq) and a 37% aqueous solution offormaldehyde (0.715 mL, 8.8 eq) were refluxed for 24 h in 5 mL ofethanol. The reaction mixture was then evaporated and the residue waswashed with pentane to yield expected compound as a pale yellow solid(0.506 g, 40% yield). LC-MS: m/z (ESI) 469.2-471.2 [M+H]⁺.

¹H NMR (300 MHz, DMSO-d₆) δ8.95; (s, 1H), 8.41; (d, J=9.0 Hz, 1H), 8.37;(d, J=5.4 Hz, 1H), 7.84; (d, J=2.2 Hz, 1H), 7.53; (dd, J=9.0 Hz and 2.0Hz, 1H), 7.08; (s, 2H), 6.59; (d, J=5.4 Hz, 1H), 3.5-3.7; (m, 12H),2.4-2.6; (m, 8H); ¹³C NMR (300 MHz, CDCl₃) δ154.3; (C), 151.3; (CH),149.7; (C), 149.0; (C), 135.5; (C), 130.0; (C), 128.3; (CH), 126.2;(CH), 125.5; (2 CH), 123.7; (2C), 121.6; (CH), 117.5; (C), 101.3; (CH),66.9; (4 CH₂), 59.1; (2 CH₂), 53.3; (4 CH₂).

Example 26 Preparation of4-[(7-chloroquinolin-4-yl)amino]-2,6-bis(4-methylpiperazin-1-ylmethyl)phenol(compound 80)

4-[(7-Chloroquinolin-4-yl)amino]phenol (of example 24—step A) (0.20 g,0.74 mmol), N-methylpiperazine (0.725 mL, 8.8 eq) and a 37% aqueoussolution of formaldehyde (0.49 mL, 8.8 eq) were refluxed for 24 h in 5mL of ethanol. The reaction mixture was then evaporated and the residuewas washed with pentane to yield expected compound as a pale yellowsolid (0.392 g, 97% yield). LC-MS: m/z (ESI) 495.3-497.3 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.39; (d, J=5.7 Hz, 1H), 8.37; (d, J=5.7 Hz,1H), 7.93; (d, J=2.1 Hz, 1H), 7.60; (dd, J=9.0 Hz and 2.1 Hz, 1H), 7.22;(s, 2H), 6.76; (d, J=5.7 Hz, 1H), 3.84; (s, 4H), 3.4-3.5; (m, 16H),2.6-2.8; (m, 16H), 2.44; (s, 6H); ¹³C NMR (300 MHz, DMSO-d₆) δ153.8;(C), 152.4; (CH), 150.0; (C), 134.3; (C), 130.7; (C), 128.0; (CH),125.1; (CH), 124.8; (CH), 123.8; (C), 118,2; (C), 100.9; (CH), 57.5;(CH₂), 54.4; (CH₂), 51.8; (CH₂), 45.2; (CH₃).

Example 27 Preparation of5-[(7-chloroquinolin-4-yl)amino]-2-(pyrrolidin-1-ylmethyl)phenol(compound 81) and5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-ylmethyl)phenol(compound 82)

Example 27.1 Preparation of5-[(7-chloroquinolin-4-yl)amino]-2-(pyrrolidin-1-ylmethyl)phenol(compound 81) Step A:N-[3-hydroxy-4-(pyrrolidin-1-ylmethyl)phenyl]acetamide

N-(3-hydroxyphenyl)acetamide (0.50 g, 3.31 mmol), formaldehyde 37% inwater (678 μL, 2.7 eq), and pyrrolidine (276 μL, 1 eq) were dissolved in10 mL of EtOH. The reaction mixture was refluxed overnight, evaporatedand purified by flash chromatography (DCM/MeOH/NH₄OH 9.6:0.4:0.1) toyield expected compound as a pale yellow oil (224 mg, 29% yield). LC-MS:m/z (ESI) 235.13 [M+H]⁺

Step B: 5-amino-2-(pyrrolidin-1-ylmethyl)phenol

N-[3-hydroxy-4-(pyrrolidin-1-ylmethyl)phenyl]acetamide (0.14 g, 0.61mmol),hydrochloric acid 20% (1 mL). The mixture was refluxed 6 h,evaporated and used for the next step.

Step C: 5-[(7-chloroquinolin-4-yl)amino]-2-(pyrrolidin-1-ylmethyl)phenol

5-amino-2-(pyrrolidin-1-ylmethyl)phenol (117 mg, 0.61 mmol) and4,7-dichloroquinoline (143 mg, 1 eq) were refluxed overnight in 1 mL ofEtOH. The reaction mixture was then evaporated and purified by flashchromatography (DCM/MeOH/NH₄OH 9:1:0.1) to yield expected compound as ayellow solid (92 mg, 29% yield). LC-MS: m/z (ESI) 354.1-356.1 [M+H]⁺

¹H NMR (300 MHz, MeOH-d₄) δ8.38; (d, J=5.6 Hz, 1H), 8.26; (d, J=9 Hz,1H), 7.85; (d, J=2 Hz, 1H), 7.49; (dd, J=9.1 and 2.1 Hz, 1H), 7.19; (d,J=8.0 Hz, 1H), 7.01; (d, J=5.7 Hz, 1H), 6.75-6.85; (m, 2H), 4.01; (s,2H), 2.85-2.95; (m, 4H), 1.9-2.0; (m, 4H)

Example 27.2 Preparation of5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-ylmethyl)phenol(compound 82) Step A: N-[3-hydroxy-4-(morpholin-4-yl)phenyl]acetamide

N-(3-hydroxyphenyl)acetamide (0.50 g, 3.31 mmol), formaldehyde 37% (678μL, 2.7 eq), and morpholine (290 μL, 1 eq) were dissolved in 10 mL ofEtOH. The reaction mixture was refluxed overnight, evaporated andpurified by flash chromatography (DCM/MeOH/NH₄OH 9.6:0.4:0.1) to yieldexpected compound as a pale yellow oil (107 mg, 13% yield). LC-MS: m/z(ESI) 251.3 [M+H]⁺; ¹H NMR (300 MHz, MeOH-d₄) δ7.09; (d, J=1.3 Hz, 1 H),6.8-6.9; (m, 2H), 3.6-3.7; (m, 4H), 3.58; (s, 2H), 2.4-2.5; (m, 4H),2.0; (s, 3H)

Step B: 5-amino-2-(morpholin-4-ylmethyl)phenol

N-[3-hydroxy-4-(morpholin-4-yl)phenyl]acetamide (0.073 g, 0.292 mmol),hydrochloric acid 20% (1 mL). The mixture was refluxed 6 h. The reactionmixture was then evaporated and purified by flash chromatography(DCM/MeOH/9.6:0.4) to yield expected compound as a pale yellow oil(0.073 mg, 22% yield) which was used directly for the next step.

Step C: 5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-ylmethyl)phenol

5-amino-2-(morpholin-4-ylmethyl)phenol (58 mg, 0.28 mmol) and4,7-dichloroquinoline (61 mg, 1.1 eq) were refluxed overnight in 1 mL ofEtOH. The reaction mixture was then evaporated and purified by flashchromatography (DCM/MeOH/NH₄OH 9:1:0.1) to yield expected compound as abrown solid (41 mg, 40% yield). LC-MS: m/z (ESI) 470.1-472.1 [M+H]⁺

¹H NMR (300 MHz, CDCl₃) δ8.56; (d, J=5.1 Hz, 1H), 8.02; (d, J=2.1 Hz,1H), 7.84; (d, J=9.0 Hz, 1H), 7.45; (dd, J=9.0 and 2.1 Hz, 1H), 7.05;(d, J=5.1 Hz, 1H), 7.01; (d, J=8.1 Hz, 1H), 6.76; (d, J=2.1 Hz, 1H),6.71; (dd, J=8.1 and 2.1 Hz, 1H), 6.56; (sl, 1H), 3.5-3.7; (m, 6H),2.5-2.6; (m, 4H).

Example 28 Preparation of3-[(7-chloroquinolin-4-yl)amino]-2-(4-methylpiperazin-1-ylmethyl)phenol(compound 83) Step A: 3-[(7-chloroquinolin-4-yl)amino]phenol

3-aminophenol (5.00 g, 25.25 mmol) and 4,7-dichloroquinoline (2.76 g, 1eq) were refluxed overnight in 5 mL of EtOH. The reaction mixture wasthen evaporated and purified by flash chromatography (DCM/MeOH/NH₄OH9:1:0.1) to yield expected compound as a pale yellow solid (6.8 g, 99%yield). LC-MS: m/z (ESI) 271.2 [M+H]⁺;

Step B:3-[(7-chloroquinolin-4-yl)amino]-2-(4-methylpiperazin-1-ylmethyl) phenol

3-[(7-chloroquinolin-4-yl)amino]phenol (0.20 g, 0.74 mmol), formaldehyde37% (62 μL, 1.1 eq), and N-methylpiperazine (90 μL, 1.1 eq) wasdissolved in 1 mL of EtOH. The mixture was refluxed overnight,evaporated and purified by flash chromatography (DCM/MeOH/NH₄OH7.5:2.5:0.2) to yield expected compound as a pale yellow oil (42 mg, 15%yield). LC-MS: m/z (ESI) 383.3-385.3 [M+H]⁺

¹H NMR (300 MHz, MeOH-d₄) δ8.36; (d, J=5.6 Hz, 1H), 8.26; (d, J=9 Hz,1H), 7.84; (d, J=2 Hz, 1H), 7.48; (dd, J=9 and 2.2 Hz, 1H), 7.11; (d,J=8.0 Hz, 1H), 6.96; (d, J=5.6 Hz, 1H), 6.7-6.8; (m, 2H), 3.75; (s, 2H),2.4-2.8; (m, 8H), 2.32; (s, 3H)

Example 29 Preparation of7-chloro-N-(3-((diethylamino)methyl)-4-fluorophenyl)quinolin-4-amine(compound 84) Step A: (2-Fluoro-5-nitro-phenyl)-methanol

To a solution of 2-fluoro-5-nitro-benzaldehyde (1000 mg, 5.91 mmoles) inMeOH (10 mL) was added NaBH₄ (182mg, 4.82 mmoles). The reaction mixturewas stirred for 3 h at 0° C. The reaction mixture was acidified with HCl1M until pH4, concentrated. 50 mL of water was added and the compoundwas extracted with Et₂O. The organic layer was dried over MgSO₄,filtered, evaporated. Expected compound was obtained as a pale yellowsolid (1007 mg, 99% yield). mp=62-64° C. ¹H NMR (300 MHz, CDCl₃) δ8.43;(dd, 6-CH, J=6.2 Hz and 2.9 Hz, 1H), 8.19; (ddd, J=9Hz, 4.5 Hz and 2.9Hz, 1H), 7.20; (dd, J=9.0 Hz and 9.0 Hz, 1H), 4.85; (d, J=3.6Hz, 2H),2.12; (s large, 1H). ¹³C NMR (75 MHz, CDCl₃) δ125.0; (CH, d, J=10.0 Hz),124.7; (CH, d, J=6.9Hz), 116.2; (CH, d, J=23.6 Hz), 58.2; (CH₂).

Step B: Toluene-4-sulfonic acid 2-fluoro-5-nitro-benzyl ester

To a solution of NaOH (0.537 g, 13.42 mmoles) in H₂O (4 mL) was added asolution of 2-fluoro-5-nitro-phenyl)-methanol (0.999 g, 5.84 mmoles) inTHF (20 mL) at 0° C. A solution of TsCl (1.892 g, 9.92 mmoles) in THF(10mL) is then added dropwise. The reaction mixture was stirred for 2h at5-10° C. 50mL of water was added and the compound was extracted withCH₂Cl₂ (3×100 mL). The organic layer was dried over MgSO₄, filtered,evaporated. Expected compound was precipited with MeOH and purified byTLC (Hex/AcOEt//7/3) and obtained as a white solid (1.700 g, 90% yield).mp=89-91° C. ¹H NMR (300 MHz, CDCl₃) δ8.18-8.26; (m, 2H), 7.82; (m, 2H),7.35; (m, 2H), 7.20; (dd, J=8.7 Hz et 8.7 Hz, 1H), 5.17; (s, 2H), 2.46;(s, 3H). ¹³C NMR (75 MHz, CDCl₃) δ130.1; (CH), 128.1; (CH), 126.7; (CH,d, J=10.0 Hz), 126.3; (CH, d, J=5.0 Hz), 116.7; (CH, d, J=23.6 Hz),64.1; (CH₂), 22.2; (CH₃).

Step C: Diethyl-(2-fluoro-5-nitro-benzyl)-amine

To a solution of toluene-4-sulfonic acid 2-fluoro-5-nitro-benzyl ester(0.696 g, 2.13 mmoles) in 1,4-dioxane (4 mL) was added TEA (0.45 mL,3.20 mmoles) and diethylamine (0.33 mL, 3.21 mmoles). The reactionmixture was stirred for 8 h at 55° C., concentrated and purified by TLC(petroleum ether/AcOEVNH₄OH//8/2/0.2). The expected compound wasobtained as a yellow oil (0.365g, 76% yield). MALDI-TOF: m/z 227.1[M+H]⁺. ¹H NMR (300 MHz, CDCl₃) δ8.44 (dd, J=6.3 Hz and 2.9 Hz, 1H),8.12; (ddd, 4-CH, J=8.9 Hz, 2.9 Hz and 4.4 Hz, 1H), 7.15; (dd, J=8.9 Hzand 8.9 Hz, 1H), 3.67; (s, 2H), 2.58; (q, J=7.1 Hz, 4H), 1.08; (t, J=7.1Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ126.7; (CH, d, J=6.9 Hz), 124.1; (CH,d, J=10.2 Hz), 115.9; (CH, d, J=24.6 Hz), 49.6; (CH₂), 47.1; (CH₂),11.8; (CH₃).

Step D: 3-((diethylamino)methyl)-4-fluorobenzenamine

To a solution of diethyl-(2-fluoro-5-nitro-benzyl)-amine (0.155 mg, 0.69mmoles) in THF (20 mL) was added a solution of SnCl₂ (520 mg, 2.74mmoles) in THF (5 mL) and HCl 1M (2.1 mL). The reaction mixture wasrefluxed for 8 h and concentrated. Saturated solution of NaHCO₃ wasadded until pH8. Aqueous layer was extracted with CH₂Cl₂ (5×50 mL). Theorganic layer was dried over MgSO₄, filtered, evaporated. Expectedcompound was precipited with MeOH and purified by TLCDCM/MeOH/NH₄OH//9/1/0.2) and obtained as a yellow oil (98 mg, 73%yield). MALDI-TOF: m/z 197.2 [M+H]⁺. ¹H NMR (300 MHz, CDCl₃) δ6.80; (dd,J=8.7 Hz and 8.7 Hz, 1H), 6.75; (dd, J=5.7 Hz and 3.0, 1H), 6.49; (ddd,J=8.7 Hz, 3.9 Hz and 3.0 Hz, 1H), 3.65-3.75; (s large, 2H), 3.57; (s,2H), 2.57; (q, J=7.2 Hz, 4H), 1.07; (t, J=7.2 Hz, 6H). ¹³C NMR (75 MHz,CDCl₃) δ118.2; (CH, d, J=3.8 Hz), 116.4; (CH, d, J=23.3 Hz), 115.7; (CH,d, J=7.9 Hz), 49.8; (CH₂), 47.3; (CH₂), 12.4; (CH₃).

Step E:7-chloro-N-(3-((diethylamino)methyl)-4-fluorophenyl)quinolin-4-amine

To a solution of 3-((diethylamino)methyl)-4-fluorobenzenamine (98.3mg,0.50 mmoles) in acetonitrile (10 mL) was added a solution of4,7-dichloroquinoleine (99.2 mg, 0.50 mmoles) in acetonitrile (5 mL) andHCl 1M (0.5 mL). The reaction mixture was refluxed overnight,concentrated and purified by TLC (DCM/MeOH/NH₄OH//9.5/0.5/0.2). Expectedcompound was obtained as a white solid (163 mg, 91% yield). MALDI-TOF:m/z 358.3-360.2 [M+H]⁺. ¹H NMR (300 MHz, CDCl₃) δ8.50; (d, J=5.4 Hz,1H), 8.00; (d, J=2.1 Hz, 1H), 7.93; (d, J=9.0 Hz, 1H), 7.47; (dd, J=6.3Hz and 2.8 Hz, 1 H), 7.42; (dd, J=9.0 Hz and 2.2 Hz, 1H), 7.21; (ddd,J=8.8 Hz, 4.5 Hz and 2.8 Hz, 1H), 7.08; (dd, J=8.8 Hz and 8.8 Hz, 1H),6.79; (d, J=5.4 Hz, 1H), 3.71; (s, 2H), 2.65; (q, J=7.2 Hz, 4H), 1.10;(t, J=7.2 Hz, 6H). ¹³C NMR (75 MHz, CDCl₃) δ151.6; (CH), 128.6; (CH),126.4; (CH, d, J=3.9 Hz), 126.1; (CH), 123.9; (CH, d, J=8.5 Hz), 121.5;(CH), 116.3; (CH, d, J=23.7 Hz), 101.9; (CH), 49.6; (CH₂), 47.0; (CH₂),11.4; (CH₃).

Example 30 Preparation of7-chloro-N-(4-fluoro-3-(pyrrolidin-1-ylmethyl)phenyl)quinolin-4-amine(compound 85) Step A: 1-(2-Fluoro-5-nitro-benzyl)-pyrrolidine

To a solution of toluene-4-sulfonic acid 2-fluoro-5-nitro-benzyl ester(compound of step B of example 29) (0.696 g, 2.31 mmoles) in 1,4-dioxane(20 mL) was added TEA (0.32 mL, 2.31 mmoles) and pyrrolidine (0.19 mL,2.31 mmoles). The reaction mixture was stirred for 24 h at roomtemperature, concentrated and purified by TLC (petroleumether/AcOEt/NH₄OH//8/2/0.2). The expected compound was obtained as ayellow oil (0.399 g, 77% yield). MALDI-TOF: m/z 225.1 [M+H]⁺. ¹H NMR(300 MHz, CDCl₃) δ8.38; (dd, J=6.2 Hz and 2.9 Hz, 1H), 8.15; (ddd, 4-CH,J=8.9 Hz, 2.9 Hz and 4.4 Hz, 1H), 7.17; (dd, J=8.9 Hz and 8.9 Hz, 1H),3.76; (s, 2H), 2.5-2.6; (m, 4H), 1.8-1.9; (m, 4H). ¹³C NMR (75 MHz,CDCl₃) δ127.0; (CH, d, J=6.4 Hz), 124.5; (CH, d, J=10.1 Hz), 116.2; (CH,d, J=24.9 Hz), 54.1; (CH₂), 52.2 (CH₂), 23.5; (CH₂).

Step B: 4-Fluoro-3-(pyrrolidin-1-ylmethyl)benzenamine

To a solution of 1-(2-fluoro-5-nitro-benzyl)-pyrrolidine (420 mg, 1.87mmol) in THF (20 mL) was added a solution of SnCl₂ (1421 mg, 7.49 mmol)in THF (5 mL) and HCl 1M (2.1 mL). The reaction mixture was refluxed for4 h and concentrated. Saturated solution of NaHCO₃ was added until pH8.Aqueous layer was extracted with CH₂O1₂ (5×50 mL). The organic layer wasdried over MgSO₄, filtered, evaporated. Expected compound was precipitedwith MeOH and purified by TLC (DCM/MeOH/NH₄OH//9.5/0.5/0.35) andobtained as a yellow oil (229.5 mg, 63% yield). MALDI-TOF: m/z 195.2[M+H]⁺. ¹H NMR (300 MHz, CDCl₃) δ6.76; (dd, J=9.5 Hz and 8.7 Hz, 1H),6.69; (dd, J=6.1 Hz and 2.9 Hz, 1H), 6.46; (ddd, J=8.7 Hz, 4.1 Hz and2.9 Hz, 1H), 3.57; (d, J=1.6 Hz, 2H), 3.3-3.5; (s large, 2H), 2.5-2.6;(m, 2H), 1.7-1.8; (m, 4H). ¹³C NMR (75 MHz, CDCl₃) δ117.5; (CH, d, J=3.5Hz), 115.7; (CH, d, J=23.6 Hz), 115.0; (CH, d, J=7.7 Hz), 54.1; (CH₂),52.8; (CH₂), 23.6; (CH₂).

Step C:7-Chloro-N-(4-fluoro-3-(pyrrolidin-1-ylmethyl)phenyl)quinolin-4-amine

To a solution of 4-fluoro-3-(pyrrolidin-1-ylmethyl)benzenamine (199.4mg, 1.03 mmoles) in acetonitrile (40 mL) was added a solution of4,7-dichloroquinoleine (203.3 mg, 1.03 mmoles) in acetonitrile (5 mL)and HCl 1M (1.12 mL). The reaction mixture was refluxed for 2 h,concentrated and purified by TLC (DCM/MeOH/NH₄OH//9/1/0.2). Expectedcompound was obtained as a white solid (330.4 mg, 90% yield). MALDI-TOF:m/z 356.2-358.1 [M+H]⁺. ¹H NMR (300 MHz, CDCl₃) δ8.36; (d, J=5.6 Hz,1H), 8.26; (d, J=9.1 Hz, 1H), 7.85; (d, J=2.1 Hz, 1H), 7.48; (dd, J=9.1Hz and 2.1 Hz, 1H), 7.40; (dd, J=6.4 Hz and 2.8 Hz, 1H), 7.31; (ddd,J=8.7 Hz, 4.5 Hz and 2.8 Hz, 1H), 7.18; (dd, J=8.8 Hz and 8.8 Hz, 1H),6.82; (d, J=5.6 Hz, 1H), 3.74; (s, 2H), 2.6-2.7; (m, 4H), 1.8-1.9; (m,4H). ¹³C NMR (75 MHz, CDCl₃) δ149.7; (CH), 125.8; (CH), 125.1; (CH),124.0; (CH), 123.5; (CH, d, J=8.8 Hz), 121.8; (CH), 114.6; (CH, d,J=23.9 Hz), 99.6; (CH), 52.1; (CH₂), 50.5; (CH₂), 21.4; (CH₂).

Example 31 Preparation of7-chloro-N-[4-(morpholin-4-yl)-3-(pyrrolidin-1-ylmethyl)phenyl]quinolin-4-amine(compound 86) Step A: [2-(Morpholin-4-yl)-5-nitrophenyl]methanol

2-Fluoro-5-nitrobenzyl alcohol (1 g, 5.84 mmol) and morpholine (1.03 mL,11.68 mmol) were heated at 115° C. for 1 h. The reaction mixture wasthen diluted with 40 mL of THF and washed with a saturated aqueoussolution of Na₂CO₃ and then with brine. The organic layer was dried overMgSO₄, filtered and evaporated to yield expected compound as yellow oil(1.38 g, 99% yield). MALDI-TOF: m/z 239.2 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.44; (d, J=2.7 Hz, 1H), 8.16; (dd, J=9.0 and2.8 Hz, 1H), 7.23; (d, J=8.7 Hz, 1H), 4.75; (s, 2H), 3.89; (t, J=4.7 Hz,4H), 3.09; (t, J=4.7 Hz, 4H); ¹³C NMR (75 MHz, MeOH-d₄) δ156.5; (C),143.4; (C), 136.6; (C), 124.1; (CH), 123.5; (CH), 118.8; (CH), 67.0;(CH₂), 59.4; (CH₂), 52.6; (CH₂).

Step B: [5-Amino-2-(morpholin-4-yl)phenyl]methanol

[2-(Morpholin-4-yl)-5-nitrophenyl]methanol (1.38 g, 5.80 mmol) washydrogenated using ammonium formate (3.67 g, 58 mmol) and Pd/C (10% Pd,0.62 g, 0.58 mmol) in 50 mL of EtOH. The mixture was stirred for 1 h atroom temperature and then filtered through a celite pad. The filtratewas evaporated and the residue was dissolved in DCM and washed with asaturated aqueous solution of Na₂CO₃. The aqueous layer was extractedwith DCM. The combined organic layers were dried over MgSO₄, filteredand evaporated to yield expected compound as a brown powder (1.21 g, 99%yield). LC-MS: m/z (ESI) 209.2 [M+H]⁺. ¹H NMR (300 MHz, MeOH-d₄) δ7.02;(d, J=15.0 Hz, 1 H), 6.83; (d, J=9.0 Hz, 1 H), 6.68 (dd, J =6.0Hz, 1H),4.71; (s, 2H), 3.82; (t, J=4.5 Hz, 4H), 2.87; (t, J=4.7 Hz, 4H).

Step C:{5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-yl)phenyl}methanolhydrochloride

[5-Amino-2-(morpholin-4-yl)phenyl]methanol (1.21 g, 5.80 mmol) and4,7-dichloroquinoline (1.38 g, 6.98 mmol) were refluxed in 10 mL ofn-pentanol for 3 h. The reaction mixture was then cooled to roomtemperature and the hydrochloride precipitate was removed by filtrationto yield expected compound as a yellow powder (1.90 g, 80% yield). LC-MS: m/z (ESI) 370.3 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.55; (d, J=9.0 Hz, 1H), 8.33; (d, J=6.9 Hz,1H), 7.93; (d, J=1.8 Hz, 1H); 7.77; (dd, J=9.0 and 2.1 Hz, 1H),7.58-7.56; (m, 1H), 7.40-7.28; (m, 2H), 6.86; (d, J=7.2 Hz, 1H), 4.78;(s, 2H), 3.85; (t, J=4.5 Hz, 4H), 2.97; (t, J=4.5 Hz, 4H).

Step D:7-chloro-N-[4-(morpholin-4-yl)-3-(pyrrolidin-1-ylmethyl)phenyl]quinolin-4-amine

To a suspension of{5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-yl)phenyl}methanolhydrochloride (0.105 g, 0.26 mmol) in 4 mL of NMP at 0° C. was addedthionyl chloride (90 μL, 1.23 mmol). The reaction mixture was warmed upto room temperature and stirred for 3 h. The reaction mixture was thenslowly added to pyrrolidine (0.65 mL, 7.75 mmol) in 1 mL of NMP at 0° C.and then stirred at room temperature for 2 h. This solution was dilutedin 100 mL of CH₂Cl₂ and washed with a saturated aqueous solution ofNaHCO₃. The organic layer was dried over MgSO₄, filtered, evaporated andpurified by flash chromatography (DCM/MeOH/NH₄OH//90/10/1) to yieldexpected compound as a yellow solid (105 mg, 96% yield). LC-MS: m/z(ESI) 423.3 [M+H]⁺. mp=187-188° C.

¹H NMR (300 MHz, CDCl₃) δ8.47; (d, J=5.4 Hz, 1H), 7.95; (d, J=2.1 Hz,1H), 7.94; (d, J=8.7 Hz, 1H), 7.40; (d, J=2.4 Hz, 1H), 7.37; (dd, J=8.7and 2.1 Hz, 1H), 7.18; (dd, J=8.4 and 2.4 Hz, 1H), 7.08; (d, J=8.4 Hz,1H), 6.83; (d, J=5.4 Hz, 1H), 3.88-3.84; (m, 4H), 3.70; (s, 2H),2.99-2.96; (m, 4H), 2.54-2.52; (m, 4H), 1.76-1.73; (m, 4H); ¹³C NMR (75MHz, CDCl₃) δ152.0; (CH), 128.8; (CH), 125.9; (2×CH), 122.9; (CH),121.9; (CH), 120.7; (CH), 102.0; (CH), 67.6; (CH₂), 55.0; (CH₂), 54.4;(CH₂), 53.4; (CH₂), 23.7; (CH₂).

Example 32 Preparation of7-chloro-N-{3-[(4-methylpiperazin-1-yl)methyl]-4-(morpholin-4-yl)phenyl}quinolin-4-amine(compound 87)

To a suspension of{5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-yl)phenyl}methanolhydrochloride (compound of step C of example 31) (0.105 g, 0.26 mmol) in4 mL of NMP at 0° C. was added thionyl chloride (90 μL, 1.23 mmol). Thereaction mixture was warmed up to room temperature and stirred for 3 h.The reaction mixture was then slowly added to 4-methylpiperazine (0.86mL, 7.75 mmol) in 1 mL of NMP at 0° C. and then stirred at roomtemperature for 2 h. This solution was diluted in 100 mL of CH₂Cl₂ andwashed with a saturated aqueous solution of NaHCO₃. The organic layerwas dried over MgSO₄, filtered, evaporated and purified by flashchromatography (Et₂O/MeOH/NH₄OH//80/20/1)to yield expected compound as ayellow solid (120 mg, 99% yield). LC-MS: m/z (ESI) 452.2 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.33; (d, J=5.7 Hz, 1H), 8.27; (d, J=9.0 Hz,1H), 7.85; (d, J=2.1 Hz, 1H), 7.50-7.42; (m, 2H), 7.30-7.2;1; (m, 2H);6.81; (d, J=5.7 Hz, 1H), 3.90-3.81; (m, 4H), 3.68; (s, 2H); 3.00-2.90;(m, 4H), 2.70-2.45; (m, 8H), 2.37; (s, 3H); ¹³C NMR (75 MHz, MeOH-d₄)δ152.0; (CH), 151.8; (C), 151.0; (C), 150.0; (C), 136.9; (C), 136.7;(C), 135.3; (C), 127.4; (CH), 127.3; (CH), 126.7; (CH), 124.9; (CH),124.8; (CH), 122.3; (CH), 119.3; (C), 120.3; (CH), 68.5; (CH₂), 57.9;(CH₂), 55.7; (CH₂), 54.6; (CH₂), 53.1; (CH₂); 28.1; (CH₃).

Example 33 Preparation ofN-{3-[(tert-butylamino)methyl]-4-(morpholin-4-yl)phenyl}-7-chloroquinolin-4-amine(compound 88)

To a suspension of{5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-yl)phenyl}methanolhydrochloride (compound of step C of example 31) (0.105 g, 0.26 mmol) in4 mL of NMP at 0° C. was added thionyl chloride (90 μL, 1.23 mmol). Thereaction mixture was warmed up to room temperature and stirred for 3 h.The reaction mixture was then slowly added to tert-butylamine (0.81 mL,7.75 mmol) in 1 mL of NMP at 0° C. and then stirred at room temperaturefor 2 h. This solution was diluted in 100 mL of CH₂Cl₂ and washed with asaturated aqueous solution of NaHCO₃. The organic layer was dried overMgSO₄, filtered, evaporated and purified by flash chromatography(Et₂O/MeOH/NH₄OH//90/10/1 then 80/20/1) to yield expected compound as ayellow solid (103 mg, 92% yield). LC-MS: m/z (ESI) 425.2 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.36; (d, J=5.6 Hz, 1H), 8.29; (d, J=9.0 Hz,1H), 7.85; (d, J=2.1 Hz, 1H), 7.49; (dd, J=9.1 and 2.4 Hz, 1H), 7.43;(d, J=1.7 Hz, 1 H), 7.38-7.30; (m, 2H); 6.91; (d, J=5.6 Hz, 1 H), 3.96;(s, 2H), 3.90-3.84; (m, 4H), 3.00-2.95; (m, 4H), 1.30; (s, 9H); ¹³C NMR(75 MHz, MeOH-d₄) δ152.5; (CH), 151.2; (C), 150.2; (C), 150.1; (C),138.0; (C), 136.8; (C), 136.;0 (C), 127.8; (CH), 127.1; (CH), 126.7;(CH), 125.3; (CH), 124.7; (CH), 123.5; (CH), 119.5; (C), 102.6; (CH),68.5; (CH₂), 54.7; (CH₂), 50.8; (C), 43.7; (CH₂), 28.1; (CH₃).

Example 34 Preparation of7-chloro-N-[3-({[2-(dimethylamino)ethyl](methyl)amino}methyl)-4-(morpholin-4-yl)phenyl]quinolin-4-amine

To a suspension of{5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-yl)phenyl}methanolhydrochloride (compound of step C of example 31) (0.100 g, 0.25 mmol) in4 mL of NMP at 0° C. was added thionyl chloride (90 μL, 1.23 mmol). Thereaction mixture was warmed up to room temperature and stirred for 3 h.The reaction mixture was then slowly added toN,N,N′-trimethylethylenediamine (1 mL, 7.50 mmol) in 1 mL of NMP at 0°C. and then stirred at room temperature for 2 h. This solution wasdiluted in 100 mL of CH₂Cl₂ and washed with a saturated aqueous solutionof NaHCO₃. The organic layer was dried over MgSO₄, filtered, evaporatedand purified by flash chromatography (DCM/MeOH/NH₄OH//90/10/1 then80/20/1) to yield expected compound as a yellow solid (97 mg, 85%yield). LC-MS: m/z (ESI) 454.4 [M+H]⁺.

¹H NMR (300 MHz, CDCl₃) δ8.50; (d, J=3.0 Hz, 1H), 8.03; (d, J=3.0 Hz,1H), 7.89; (d, J=9 Hz, 1H), 7.42; (m, 2H), 7.26; (m, 2H), 6.84; (d,J=3.0 Hz, 1H), 3.88-3.85; (m, 4H), 3.68; (s, 2H), 3.00-2.97; (m, 4H),2.52-2.49; (m, 4H) ¹³C NMR (75 MHz, CDCl₃-d₁) δ152.4; (C), 151.9; (CH),149.8; (CH), 148.4; (CH), 135.7; (CH), 135.5; (CH), 135.3; (CH), 128.9;(C), 126.0; (C), 125.7; (C), 122.3; (C), 120.6; (C), 118.2; (CH₂),101.8; (C), 67.6; (CH₂), 57.2; (CH₂), 54.7; (CH₂), 53.5; (CH₃)

Example 35 Preparation of7-chloro-N-[4-(morpholin-4-yl)-3-(morpholin-4-ylmethyl)phenyl]quinolin-4-amine(compound 90)

To a suspension of{5-[(7-chloroquinolin-4-yl)amino]-2-(morpholin-4-yl)phenyl}methanolhydrochloride (compound of step C of example 31) (0.100 g, 0.25 mmol) in4 mL of NMP at 0° C. was added thionyl chloride (90 μL, 1.23 mmol). Thereaction mixture was warmed up to room temperature and stirred for 3 h.The reaction mixture was then slowly added toN,N,N′-trimethylethylenediamine (0.65 mL, 7.38 mmol) in 1 mL of NMP at0° C. and then stirred at room temperature for 2 h. This solution wasdiluted in 100 mL of CH₂Cl₂ and washed with a saturated aqueous solutionof NaHCO₃. The organic layer was dried over MgSO₄, filtered, evaporatedand purified by flash chromatography (DCM/MeOH///90/10) to yieldexpected compound as a yellow solid (83 mg, 77% yield). LC-MS: m/z (ESI)439.3 [M+H]⁺.

¹H NMR (300 MHz, MeOH-d₄) δ8.50; (d, J=3.0 Hz, 1H), 8.03; (d, J=3.0 Hz,1H), 7.89; (d, J=9.0 Hz, 1H), 7.42; (m, 2H), 7.26; (m, 2H), 6.84; (d,J=3.0 Hz, 1H), 3.88-3.85; (m, 4H), 3.82-3.70; (m, 4H), 3.69; (s, 2H),3.00-2.97; (m, 4H), 2.52-2.49; (m, 4H), 2.10-1.92; (m, 5H)¹³C NMR (75MHz, MeOH-d₄) δ149.7; (CH), 148.6; (CH), 135.7; (CH), 134.9; (CH),134.7; (CH), 128.7; (C), 126.3; (C), 125.8; (C), 123.0; (C), 121.4; (C),117.9; (CH), 67.6; (CH₂), 67.2; (CH₂), 57.9; (CH₂), 53.9; (CH₂), 53.5;(CH₂).

B—Biological Tests In vitro Experiments Cell treatment for Cytotoxicityand Western Blotting

The human neuroblastoma cell line SKNSH-SY5Y APP^(wt) is cultured inDulbecco's modified Eagle medium (Invitrogen) supplemented with 10%fetal calf serum (PAA), 2 mM L-glutamine (Invitrogen), 1 mMnon-essential amino acids (Invitrogen), 50 units/mlpenicillin/streptomycin (Invitrogen) and 200 μg geneticin (G418;Invitrogen) (selection for cell expressing APP) in a 5% CO₂ humidifiedincubator at 37° C. Cells are seeded into 96-wells plates (20000cells/well) for cytotoxicity assay and 12-wells plate (300000cells/well) for western blotting for 18 h.

Compounds are dissolved in pure DMSO at a concentration of 10 mM (StockSolution) over a silica gel layer under a chemical hood. Forcytotoxicity assay, cells are incubated with 100 μl/well of each productat 100; 30; 10; 3; 1; 0.3 and 0.1 μM in a 5% CO₂ incubator at 37° C. for24 h. The number of viable cells is determined with CellTiter 96®Aqueous One solution Reagent (MTS) (Promega) according to themanufacturer protocol. Briefly, 20 μl of this reagent is added to eachwell and the plate is incubated for 2 hours in a 5% CO₂ incubator at 37°C. The absorbance is read at 490 nm. Optical density is analyzed withexcel computer program and CC50 determined with Graph Pad Prism (version4.02) computer program.

For western blotting, cells are incubated with 700 μl/well of eachproduct at 10; 3; 1 and 0.31,1M in a 5% CO₂ incubator at 37° C. for 24h. The following day, culture medium samples are collected for ELISAassay and stored at −80° C. Cells are rinsed very slowly with cold PBS,snap-frozen on dry ice and immediately stored at −80° C.

Innotest β Amyloid 1-42 Innogenetics®

The β-amyloid 1-42 rate is quantified in SY5Y cells-culture medium byusing the Innotest β-amyloid 1-42 (80324 Innogenetics®) according to themanufacturer instructions. Briefly, Fresh culture medium is used todilute the standard. Culture samples are centrifuged at 200 g at 4° C.during 5 minutes in order to eliminate cell fragments.

First, 75 μl of the conjugate working solution1 (3D6 biotinylatedantibody) is added to a 21F12 Ab precoated well. Twenty five μl of puresupernatant samples are then added to the coated well, and incubated for1 h. Then the plate is washed 5 times with the washing buffer. Then 100μl of peroxidase-labeled streptavidin (conjugate working solution 2) isadded and incubated for 30 min at RT. After washing the plate 5 times,100 μl of tetramethyl benzidine (substrate working solution) is addedonto the plate and incubated for 30 minutes in the dark. The reaction isstopped with 50 μl 0.9N sulfuric acid (stop solution). The absorbance isread at 450 nm.

A sigmoidal curve fitting (four parameter polynomial curve) is used tocalculate the standard curve. The corresponding concentration ofβ-amyloid (1-42) in pg/ml is determined from the standard curve. Resultsare expressed in concentration of β-amyloid in pg per μg of totalproteins. Graphs and statistic analyses (t Test) are performed withGraph Pad Prism computer program (V0.5).

In Vivo Experiments Mice Treatment

Test molecules are intraperitoneally injected to 4 month old C57BI6females mice either at 12.5; 25; 50 mg/kg or vehicule once, andsacrified 24 h after by decapitation. Brain is immediately removed andPrefrontal Cortex is dissected by using a cold zinc brain matrix. Thetotal dissection time was <3 min from decapitation. Following dissectiontissue is frozen in dry ice and then stored at −80° C. for westernblotting.

Protein Extraction and Western Blotting Analysis

Cells in twelve-well plates are collected in 50 μl Laemmli Lysis Buffer(pH 6.8, 50 mM Tris Base, 2% SDS, 20% Glycerol, 1 mM sodiumpyrophosphate, 1 mM sodium orthovanadate and Protease Inhibitor CocktailComplete Mini-Roche) per well. Cells are scrapped and lysates sonicatedfor 5 min.

Protein extracts of Prefrontal Cortex samples are prepared in 50 μlLaemmli Lysis Buffer (pH 6.8, 50 mM Tris Base, 2% SDS, 20% Glycerol, 1mM sodium pyrophosphate, 1 mM sodium orthovanadate and ProteaseInhibitor Cocktail Complete Mini-Roche) by trituration of the fragmentsin a 0.2 ml glass micro tissue grinder. The tissue lysates are sonicatedfor 5 min, then passed through a 26 gauge needle and rocked for 1 h at4° C. Samples are then cleared by centrifugation at 200 g for 5 min.

Protein content is determined using the BCA Method (Pierce). Samplescontaining equal amounts of protein (35 μg total protein for mice and 20μg total protein for cells per well) are heated at 85° C. for 2 minafter thawing and electrophoresed for 2.45 h at 125 V in a 16% Tricineprecast gels according to the protocol supplied with the NuPAGE system(Invitrogen). After size fractionation, the proteins are transferredonto nitrocellulose 0.2 μm pore-size membranes (Invitrogen) in the blotmodule of the NuPAGE sytem for 1 h at room temperature (RT). Blots areblocked for 1 h in TNT with 0.05% Tween 20 and 5% nonfat milk at RT,incubated overnight at 4° C. with anti-C term APP C1:3-2429 (1:250000)primary antibody and washed four times with TNT before being exposed toperoxidase labeled anti-mouse IgG (H+L) C4 94010 (1:50000; Vector)secondary antibody for 45 h at 25° C. The immunoreactions are detectedwith SuperSignal West Pico Chemiluminescent Substrate (Thermo specific,34077). Images are scanned with LAS-3000 Image System (Global FUJIFILM)and bands quantified by ImageJ 1.37v computer program. The differentAPP-CTF bands are detected as a function of their molecular mass, asdescribed in Vingtdeux, V., Hamdane, M., Gompel, M. et al. (2005)Phosphorylation of amyloid precursor carboxy-terminal fragments enhancestheir processing by a gamma-secretase-dependent mechanism. NeurobiolDis, 20, 625-637. Graphs and statistical analysis of results isperformed with Graph Pad Prism (version 4.02) computer program.

The full biological results are given below (for known compounds as wallas for new compounds of examples 1 to 23 as mentioned above) (see alsoFIG. 1 for in vivo results).

Biological Results of Compounds having Formula (I-4-1)

CC₅₀ μM CTFalpha AICD MRC-5 N^(o) Ref Ref lab R₆ NR₁R₂ 1 μM* 1 μM*(SY5Y) 1 EP-29 EP985 H NEt₂ 1119 413 31.3 ± 0.8 3 EP-23 EP637 OEt NEt₂3071 129 — 4 EP-30 EP537 H pyrroli- 1065 378 29.8 ± 6.2 dine *results inarbitrary units compared to a control with a value of 100

Biological Results of Compounds having Formula (I-4-2)

CTFα AICD CC₅₀ μM N^(o) Ref Ref lab R₆ NR₁R₂ 1 μM* 1 μM* MRC-5 13 EPC-EP987 pF—C₆H₄— NEt₂ 1560 176 17.4 ± 0.2 9g $ 19 EPC- EP671 Et NEt₂ 1349194 18.1 ± 1.2 9m $ 20 EPC- EP1031 pCF₃O—C₆H₄— NEt₂ 1880 150 18.6 ± 0.89n $ 645 34 EPC- EP517 Me pyrrolidine 1040 332 21.3 ± 3.4 10l $ 35 EPC-EP541, Et pyrrolidine 1665 328 18.9 ± 3.8 10m 441 *results in arbitraryunits compared to a control with a value of 100

Biological Results of Compounds having Formula (I-4-3)

Aβ CTFα AICD CC₅₀ μM N^(o) Ref Ref lab NR_(a)R_(b) NR₁R₂ 1 μM* 1 μM* 1μM* MRC-5 42 EPN- EP573 morpholine NEt₂ 90 155 189 16.7 ± 0.2 7d 47 EPN-EP977 dimethylamine pyrrolidine 15 109 130 14.7 ± 1.2 8a 50 EPN- EP577morpholine pyrrolidine 55 311 201 31.8 ± 0.9 8d 51 EPN- EP867N-methyl-piperazine pyrrolidine 47 3569 1295  8.6 ± 0.8 8e (<5) 54 EPN-EP1101 piperidinethylamine pyrrolidine 30 1422 343  2.1 ± 0.1 8h*results in arbitrary units compared to a control with a value of 100

Biological Results of Compounds having Formula (I-5)

Aβ CTFα N^(o) Ref Ref lab R₅ NR_(a)R_(b) NR₁R₂ 1 μM* 1 μM* AICD 1 μM* 5612b NLF54 H N—Me NHtBu 45 183 823 piperazine 57 15b NLF72 H N—MeMorpholine 19 194 757 piperazine 59 23a NLF45 CH₃ N—Me Pyrrolidine 73629 325 piperazine *results in arbitrary units compared to a controlwith a value of 100

Biological Results of Compounds having Formula (I-6)

Aβ CTFα AICD N^(o) Ref Ref lab X R₅ NR₁R₂ 1 μM* 1 μM* 1 μM* 64 2cEB344(2) CO H dimethyl[3-(piperazin- 31 106 669 1-yl)propyl]amine 66 8FD15 CH₂ OH Pyrrolidine 19 754 886 *results in arbitrary units comparedto a control with a value of 100

Biological Results of Compounds having Formula (I-7)

Aβ CTFα AICD N^(o) Ref Ref lab NR₁R₂ 1 μM* 1 μM* 1 μM* 69 3c EB344(1)dimethyl[3- 46 126 302 (piperazin- 1-yl)propyl]amine *results inarbitrary units compared to a control with a value of 100

Biological Results of Compounds having Formula (I-8)

Ref Aβ CTFα AICD N^(o) Ref lab R₆ NR₁R₂ 1 μM* 1 μM* 1 μM* 70 20a EB551N—Me N—Me — 687 515 piperazine piperazine 72 21 EB546 OH N—Me 100 105 80piperazine *results in arbitrary units compared to a control with avalue of 100

Biological Results of Compounds having Formula (I-13)

Aβ CTFα AICD N^(o) Ref Ref lab structure 1 μM* 1 μM* 1 μM* 74 3 EB358a

44 4016 1933 76 5 EB418

76 2618 1503 77 7 EB374

85 1868 618 *results in arbitrary units compared to a control with avalue of 100

Biological Results of Compounds having Formula (I-9)

CC₅₀ Aβ CTFα AICD μM N^(o) Ref Ref lab NR₁R₂ 1 μM* 1 μM* 1 μM* MRC-5 8040 MAD1342 NMe(CH₂)₂ NMe₂ 55 570 1226 <10 83 43 SD5/4 NH(CH₂)₂Nmorpholine 80 286 135 <10 84 44 SD4/54 NEt₂ 60 358 209 <10 86 47 SD4/35N(pyrrolidine) 30 856 664 <10 87 48 SD4/14 N(piperazine)N—Me 40 1019 255<10 91 52 SD4/36 N(piperazine)N—Ph 25 884 286 <10 95 56 SD4/73N(piperazine)N—CH₂—pOMePh 45 693 191 <10 101 62 MAD1338 NHCH₂Ph 20 315329 <10 103 64 SD4/78 NHCH₂—pCIPh 20 186 133 <10 *results in arbitraryunits compared to a control with a value of 100

Biological Results of Compounds having Formula (I-10)

Aβ CTFα AICD N^(o) Ref Ref lab NR₁R₂ 1 μM* 1 μM* 1 μM* 107 5 SD5/56N(morpholine) 25 1013 450 *results in arbitrary units compared to acontrol with a value of 100

Biological Results of Compounds having Formula (I-10)

Aβ CTFα AICD CC₅₀ μM N^(o) Ref Ref lab NR₁R₂ 1 μM* 1 μM* 1 μM* MRC-5 10910 MAD1328 NHCH₂CH₂CH₂NMe₂ 75 100 100 >32 110 12 MAD1420NHCH₂CH₂Npyrrolidine 80 131 143 17 *results in arbitrary units comparedto a control with a value of 100

Biological Results of Compounds having Formula (I-12)

Aβ CTFα AICD CC₅₀ μM N^(o) Ref Ref lab NR₁R₂ 1 μM* 1 μM* 1 μM* MRC-5 1132 SD4/18 NH (CH₂)₂ N-morpholine 80 77 170 >12.5 114 3 SD3/34NH(CH₂)₃NEt₂ 70 240 203 6 *results in arbitrary units compared to acontrol with a value of 100

In vivo Results of Compound 66 (See Example 12)

FIG. 1 represents the evolution of CTF-alpha and CTF-beta fragments byC57B16 female mice to which either 12.5, 25, 50 mg/kg of compound 66 oronly the vehicle was injected (control). The results show the opticaldensity (in arbitrary units) for each mouse. FIG. 1 shows that compound66 increases the production of APP-CTFα.

1-13. (canceled)
 14. A compound of formula (I-5):

wherein R₁ and R₂ are each independently chosen from: H, alkyl, aralkyl,aryl, cycloalkyl and heterocycle groups, said alkyl, aralkyl, aryl,cycloalkyl and heterocycle groups being substituted or unsubstituted, orR₁ and R₂ may form together with the nitrogen atom carrying them asubstituted or unsubstituted heterocycle group; R_(a) and R_(b) are eachindependently chosen from: H, alkyl, aralkyl, aryl, cycloalkyl, andheterocycle groups, said alkyl, aralkyl, aryl, cycloalkyl, andheterocycle groups being substituted or unsubstituted, or R_(a) andR_(b) may form together with the nitrogen atom carrying them asubstituted or unsubstituted heterocycle group; R₅ is chosen from thecroup consisting of: H, (C₁-C₁₂)alkyl, OH, and (C₁-C₁₂)alkoxy, or itspharmaceutically acceptable salts, hydrates or hydrated salts or itspolymorphic crystalline structures, racemates, diastereisomers orenantiomers.
 15. A compound of formula (I-6):

wherein R₁ and R₂ are each independently chosen from: H, alkyl, aralkyl,aryl, cycloalkyl and heterocycle groups, said alkyl, aralkyl, aryl,cycloalkyl and heterocycle groups being substituted, or unsubstituted,or R₁ and R₂ may form together with the nitrogen atom carrying them asubstituted or unsubstituted heterocycle group, X is CO or CH₂ and R₅ isH.
 16. A compound of formula (I-7):

wherein R₁ and R₂ are each independently chosen from: H, alkyl, aralkyl,aryl, cycloalkyl and heterocycle groups, said alkyl, aralkyl, aryl,cycloalkyl and heterocycle groups being substituted or unsubstituted, orR₁ and R₂ may form together with the nitrogen atom carrying them asubstituted or unsubstituted heterocycle group.
 17. A compound offormula (I-8):

wherein R₁ and R₂ are each independently chosen from: H, alkyl, aralkyl,aryl, cycloalkyl and heterocycle groups, said alkyl, aralkyl, aryl,cycloalkyl and heterocycle groups being substituted or unsubstituted,and wherein R₁ and R₂ form together with the nitrogen atom carrying thema heterocycle, and R₆ is chosen from the group consisting of: H,halogen, CN, OH, (C₁-C₁₂)alkyl, heteroaryl, CO₂R, wherein R is an alkylgroup comprising from 1 to 12 carbon atoms. NR_(a)R_(b), R_(a) and R_(b)being each independently chosen from: H, alkyl, aralkyl, aryl,cycloalkyl, and heterocycle groups, said alkyl, aralkyl, aryl,cycloalkyl, and heterocycle groups being substituted or unsubstituted,or R_(a) and R_(b) forming together with the nitrogen atom carrying thema substituted or unsubstituted heterocycle group; and a radical offormula: —X′—CH₂)^(n)—NR′₁R′₂, wherein: X′ is chosen from: CH₂, O, NH,CO, CH₂OCO, and NHCO; n′ is 0, 1 or 2; R′₁ and R′₂ are eachindependently chosen from H, alkyl, aralkyl. aryl, cycloalkyl andheterocycle groups, said alkyl, aralkyl, aryl, cycloalkyl, andheterocycle groups being substituted or unsubstituted, or R′₁ and R′_(z)may form together with the nitrogen atom carrying them a substituted orunsubstituted heterocycle group.
 18. A pharmaceutical compositioncomprising the compound according to claim 14, in association with apharmaceutically acceptable vehicle.
 19. A method of preventing and/ortreating a disease involving formation of amyloid plaques and/or where adysfunction of the APP metabolism occurs in a patient in need thereof,comprising the step of administering to said patient a therapeuticallyeffective amount of a compound having formula (I):

wherein: a represents a single bond or a double bond; b represents asingle pond or a double bond, provided that when a is a single bond,then b is a double bond, and when a is a-double bond, then b is a singlebond; R₃ is absent when a is a double bond, or, when a is a single bond,R₃ is chosen from the groups consisting of: alkyl, cycloalkyl, aryl andheterocycle radicals, said alkyl, cycloalkyl, aryl and heterocycleradicals being substituted or unsubstituted; R₄ is absent when b is adouble bond, or, R₄ is H when b is a single bond; R₅ is chosen from thegroup consisting of: H, (C₁-C₁₂)alkyl, OH, and (C₁-C₁₂)alkoxy, R₆ ischosen from the group consisting of H, halogen, CN, OH, (C₁-C₁₂)alkoxy,(C₁-C₁₂)alkyl, (C₆-C₃)aryl, heteroaryl, CO₂R, wherein R is an alkylgroup comprising from 1 to 12 carbon atoms, NR₃R_(b), R_(a) and R_(b)being each independently chosen from: H, alkyl, aralkyl, aryl,cycloalkyl, and heterocycle groups, said alkyl, aralkyl, aryl,cycloalkyl, and heterocycle groups being substituted or unsubstituted,or R_(a) and R_(b) forming together with the nitrogen atom carrying thema substituted or unsubstituted heterocycle group; and a radical offormula: —X′—(CH₂)_(n)—NR′₁R′₂, wherein: X′ is chosen from: CH₂, O, NH,CO, CH₂OCO, and NHCO; n′ is 0, 1 or 2; R′₁ and R′₂ are eachindependently chosen from H, alkyl, aralkyl, aryl, cycloalkyl andheterocycle groups, said alkyl, aralkyl, aryl, cycloalkyl, andheterocycle groups being substituted or unsubstituted, or R′₁ and R′₂may form together with the nitrogen atom carrying them a substituted orunsubstituted heterocycle group; X is chosen from: CH₂, O, NH, CO,CH₂OCO, and NHCO; n 0, 1 or 2; R₁ and R2 are each independently chosenfrom H, alkyl, aralkyl, aryl, cycloalkyl and heterocycle groups, saidalkyl, aralkyl, aryl, cycloalkyl and heterocycle groups beingsubstituted or unsubstituted, or R₁ and R₂ may form together with thenitrogen atom carrying them a substituted or unsubstituted heterocyclegroup; or its pharmaceutically acceptable salts, hydrates or hydratedsalts or its polymorphic crystalline structures, racemates,diastereisomers or enantiomers, with the exclusion of the compoundshaving the following formula:

wherein: R₆ is H and R₈ is —CH₂—NEt₂, R₆ is OH and R₈ is chosen from:—CH₂—NEt₂, —CH₂—NHEt,

and R₆ is OMe and R₈ is —CH₂—NEt₂ or


20. The method of claim 19, wherein said compound has formula (I-1):

wherein R₁, R₂, R₅, R₆, X, and n are as defined in claim
 19. 21. Themethod of claim 19, wherein said compound has formula (I-2):

wherein R₁, R₂, R_(5,) R₆, X, and n are as defined in claim
 19. 22. Themethod of claim. 19, wherein said compound has formula (I-3):

wherein R₁, R₂, R₅, and R₆ are as defined in claim
 19. 23. The method ofclaim 19, wherein said compound has formula (I-4):

wherein R₁ and R₂ are as defined in claim 19, and R₆ is chosen from: H,halogen, OH, (C₁-C₁₂)alkoxy, (C₁-C₁₂)alkyl, (C₆-C₃₀)aryl, hetetoaryl,and NR_(a)R_(b), and R_(a) and R_(b) are as defined in claim
 19. 24. Themethod of claim 19, wherein said compound has formula (I-5):

wherein R₁, R₂, R₅, R_(a) and R_(b) are as defined in claim 19,
 25. Themethod of claim 19, wherein said compound has formula (I-6):

wherein R₁, R₂ and R₅ are as defined in claim 19, and wherein X is CO orCH₂.
 26. The method of claim 19, wherein said compound has formula(I-7):

wherein R₁ and R₂ are as defined in claim
 19. 27. The method of claim19, wherein said compound has formula (I-8):

wherein R₁ and R₂ are as defined in claim 19, and form, together withthe nitrogen. atom carrying them, a heterocycle, and R₆ is as defined inclaim
 19. 28. The method of claim 19, wherein said compound has formula(I-9):

wherein R₁ and R₂ are as defined in claim
 19. 29. The method of claim19, wherein said compound has formula (I-10):

wherein R₁ and R₂ are as defined in claim
 19. 30. The method of claim19, wherein said compound has formula (I-11):

wherein R₁ and R₂ are as defined in claim
 19. 31. The method of claim19, wherein said compound has formula (I-12):

wherein R₁ and R₂ are as defined in claim
 19. 32. The compound of claim14, wherein R₅ is H or CH₃.
 33. The compound of claim 14, wherein R_(a)and R_(b) form, with the nitrogen atom carrying them, a substituted orunsubstituted heterocycle group.
 34. The compound of claim 33, whereinR_(a) and R_(b) form with the nitrogen. atom carrying them aN-methylpiperazine or a morpholine group.
 35. The method of claim 24,wherein R₅ is H or CH₃.
 36. The method of claim 24, wherein R_(a) andR_(b) form, with the nitrogen atom carrying them, a substituted orunsubstituted heterocycle group.
 37. The method of claim 36, whereinR_(a) and R_(b) form with the nitrogen atom carrying them aN-methylpiperazine or a morpholine group.
 38. The method of claim 25,wherein R₅ is H or OH.
 39. A pharmaceutical composition comprising thecompound according to claim 15, in association with a pharmaceuticallyacceptable vehicle.
 40. A pharmaceutical composition comprising thecompound according to claim 16, in association with a pharmaceuticallyacceptable vehicle.
 41. A pharmaceutical composition comprising thecompound according to claim 17, in association with a pharmaceuticallyacceptable vehicle,
 42. The compound of claim 17, wherein said halogenis F or Br.
 43. The method of claim 19, wherein said halogen is F or Br.